Methods and compositions associated with the granulocyte colony-stimulating factor receptor

ABSTRACT

Some embodiments include compositions and methods of using or identifying compounds that modulate the activity of the granulocyte colony-stimulating factor receptor (GCFR). Some embodiments include use of compounds to treat certain disorders, such as hematopoietic or neurological disorders.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.15/247,164 filed Aug. 25, 2016 which is a continuation of U.S.application Ser. No. 14/357,551 filed May 9, 2014, now U.S. Pat. No.9,492,430 issued Nov. 15, 2016 which is the U.S. National Phase of App.No. PCT/US2012/064706 entitled “METHODS AND COMPOSITIONS ASSOCIATED WITHTHE GRANULOCYTE COLONY-STIMULATING FACTOR RECEPTOR” filed Nov. 12, 2012and published in English on May 23, 2013 as WO 2013/074459 which claimsthe benefit of U.S. Prov. App. No. 61/561510 filed Nov. 18, 2011, andU.S. Prov. App. No. 61/559660 filed Nov. 14, 2011, each entitled“METHODS AND COMPOSITIONS ASSOCIATED WITH THE GRANULOCYTECOLONY-STIMULATING FACTOR RECEPTOR” the contents of which are herebyincorporated by reference in their entireties.

REFERENCE TO SEQUENCE LISTING

The present application is being filed along with a Sequence Listing inelectronic format. The Sequence Listing is provided as a file entitledLIGAND200C2SEQ.TXT, created Aug. 28, 2018, which is approximately 3 Kbin size. The information in the electronic format of the SequenceListing is incorporated herein by reference in its entirety.

FIELD OF THE INVENTION

Compositions and methods in the fields of chemistry and medicine aredisclosed. Some of the disclosed embodiments are compositions thatmodulate the activity of the granulocyte colony-stimulating factorreceptor (GCFR). Some of the disclosed embodiments are and methods ofusing or identifying compounds that modulate the activity of thegranulocyte colony-stimulating factor receptor (GCFR). Some of thedisclosed embodiments include the use of compounds to treat certaindisorders, such as hematopoietic or neurological disorders, associatedwith the granulocyte colony-stimulating factor receptor (GCFR).

BACKGROUND OF THE INVENTION

Granulocyte colony-stimulating factor (GCSF) is one of the hematopoieticgrowth factors with multifunctional activities. As a glycoprotein, GCSFplays important regulatory functions in the processes of maturation,proliferation, differentiation, and functional activation of granulocyteprecursors and mature granulocytes in the bone marrow. It is able toaugment white blood cell production when bone marrow dysfunction exists.Recombinant DNA technology has made it possible to clone the genesresponsible for GCSF and to develop pharmaceutical products to treat anumber of human hematopoietic conditions and disorders such asneutropenia and hematopoietic stem cell transplantation.

Human GCSF (hGCSF) protein has a molecular mass of 19.6 kDa and exertsits biological functions through binding to the human GCSF receptor(hGCSFR), a single transmembrane protein with a large extracellularregion that consists of an immunoglobulin-like (Ig-like) domain, acytokine receptor homology (CRH) domain, and three fibronectin type IIIdomains. Binding of GCSF to the extracellular Ig-like and CRH domains ofthe receptor triggers receptor homodimerization with a 2:2 stoichiometryof hGCSF/hGCSFR (Tamada T, et al. 2006 Homodimeric cross-over structureof the human GCSF receptor signaling complex. PNAS 103:3135-3140). Thedimerization results in activation of intracellular Janus tyrosinekinase-signal transducers and activators of transcription (Jak-Stat)type signaling cascade. The signaling transfer of hematopoietic factorreceptors from extracellular region to intracellular cascades has beensuggested to be via conformational changes of the receptor dimer in TMdomains. It has been demonstrated that the dimeric erythropoietin (EPO)receptor can be activated by mutations at the TM domain in the absenceof the natural ligand EPO, a hematopoietic growth factor regulating redblood cell production (Lu X, et al. 2006 Active conformation of theerythropoietin receptor: Random and cysteine-scanning mutagenesis of theextracellular juxtamembrane and transmembrane domains. JBC281:7002-7011). Patients with mutations in TM domain of hGCSFR haveexperienced chronic neutrophilia due to the receptor constitutiveactivation (Plo I, et al. 2009 An activating mutation in the CSF3R geneinduces a hereditary chronic neutrophilia. JEM 206:1701-1707).

Development of bioengineered EPO and GCSF targeted binding sites of thenatural ligands have been successful, while development of aprotein-based thrombopoietin (TPO) drug has failed due to drug-relatedincrease in antibody production against endogenous TPO. A peptide-basedTPO mimic molecule, romiplostim, recently received FDA approval,although the risk of drug-related antibody development is to bemonitored in longer term use. Discovery efforts to find small moleculestargeting the same site have not been fruitful. Eltrombopag is the firstFDA approved small molecule drug in the hematopoietic growth factorfield for the treatment of thrombocytopenia. Different from romiplostimthat competes with endogenous TPO for the same binding site of TPOreceptor, eltrombopag activates TPO receptor most likely by interactingwith the TM domain and, as a result, its activity is additive to that ofendogenous TPO. Eltrombopag showed a unique species-specific TPOreceptor activation that requires histidine-499, and partially activateda mouse GCSF receptor where a cysteine residue in the TM domain wasmutated to histidine (Erickson-Miller C, et al. 2004 Species specificityand receptor domain interaction of small molecule TPO receptor agonists.ASH 2004 San Diego, poster). Several classes of compounds have beenreported to activate the TPO receptor. For example, WO 2004/033433, WO2007/062078, WO 2006/047344, WO 01/89457, WO 2009/103218, US 7,026,334,WO 2005/014561, WO 2005/007651, WO 2006/033005, WO 2007/004038, WO2007/036769, and WO 2007/054783.

A small molecule that selectively activates mouse GCSF receptor has beenidentified. (Tian S-S, et al. 1998 A small, nonpeptidyl mimics ofgranulocyte-colony-stimulating factor. Science 281:257-259). Thismolecule is reported to not directly compete with GCSF, although itseems to bind to the extracellular region of mouse GCSF receptor. (DoyleML, et al. 2003 Selective binding and oligomerization of the murine GCSFreceptor by a low molecular weight, nonpeptidyl ligand. JBC278:9426-9434). A class of small molecules have been reported toactivate the both mouse and human GCSF receptors and the site of actionis not suggested (Kusano K, et al. 2004 A potential therapeutic role forsmall nonpeptidyl compounds that mimic human GCSF. Blood 103:836-842;Tokizawa M, et al. 2004 Imidazole derivatives of their salts. U.S. Pat.No. 6,737,434).

SUMMARY OF THE INVENTION

Compositions that modulate the activity of the granulocytecolony-stimulating factor receptor (GCFR) are disclosed. Methods ofusing and methods of identifying compounds that modulate the activity ofthe granulocyte colony-stimulating factor receptor (GCFR) are disclosed.Other embodiments are disclosed that include use of compounds to treatcertain disorders, such as hematopoietic or neurological disorders.

Some embodiments include a compound of Formula (I), (II), (III), or(IV):

a tautomer thereof, or a pharmaceutically acceptable salt thereof,wherein:

R¹ is selected from hydrogen, OR⁶, NO₂, CN, NR⁶R⁷, CO₂R⁶, C(═O)NR⁶R⁷,SO₃R⁶, SO₂NR⁶R⁸, an optionally substituted C₁-C₆ alkyl, an optionallysubstituted C₂-C₆ alkenyl, an optionally substituted C₂-C₆ alkynyl, anoptionally substituted C₁-C₆ heteroalkyl, an optionally substitutedC₃-C₆ cycloalkyl, an optionally substituted C₃-C₆ cycloalkenyl, anoptionally substituted C₂-C₆ heterocyclyl, an optionally substitutedarylalkyl, an optionally substituted aryl, and an optionally substitutedheteroaryl;

R² and R³ are independently selected from hydrogen, an optionallysubstituted C₁-C₆ alkyl, an optionally substituted C₂-C₆ alkenyl, anoptionally substituted C₂-C₆ alkynyl, an optionally substituted C₁-C₆heteroalkyl, an optionally substituted C₃-C₈ cycloalkyl, an optionallysubstituted C₃-C₈ cycloalkenyl, an optionally substituted C₁-C₆heterocycle, an optionally substituted aryl, and an optionallysubstituted heteroaryl;

R⁴ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₂-C₆ alkenyl, an optionally substituted C₂-C₆alkynyl, an optionally substituted C₁-C₆ heteroalkyl, an optionallysubstituted C₃-C₈ cycloalkyl, an optionally substituted C₃-C₈cycloalkenyl, an optionally substituted C₁-C₆ heterocycle, an optionallysubstituted aryl, an optionally substituted heteroaryl, an optionallysubstituted arylalkyl, an optionally substituted arylalkenyl, anoptionally substituted arylalkynyl, and an optionally substitutedheteroarylalkyl;

R⁵ is selected from hydrogen, halogen, NO₂, CN, CF₃, OR⁶, CO₂R⁶,C(═O)NR⁶R⁷, SO₃R⁶, and SO₂NR⁶R⁸, an optionally substituted aryl, anoptionally substituted C₁-C₆ alkyl, and an optionally substituted C₁-C₆heteroalkyl;

R⁶ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, aC₁-C₆ heteroalkyl, an optionally substituted aryl, and an optionallysubstituted heteroaryl;

R⁷ is selected from hydrogen, C(═O)R⁸, C(═O)NHR⁸, an optionallysubstituted C₁-C₆ alkyl, and an optionally substituted C₁-C₆heteroalkyl; or —NR⁶R⁷ is an optionally substituted non-aromaticheterocycle linked through a ring nitrogen;

R⁸ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, andan optionally substituted C₁-C₆ heteroalkyl;

R⁹ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₂-C₆ alkenyl, an optionally substituted C₂-C₆alkynyl, an optionally substituted C₁-C₆ heteroalkyl, an optionallysubstituted C₃-C₈ cycloalkyl, an optionally substituted C₃-C₈cycloalkenyl, an optionally substituted C₁-C₆ heterocycle, an optionallysubstituted heteroaryl, an optionally substituted arylalkyl, anoptionally substituted arylalkenyl, an optionally substitutedarylalkynyl, an optionally substituted heteroarylalkyl, an optionallysubstituted heteroarylalkenyl, and an optionally substitutedheteroarylalkynyl;

Q is selected from the group consisting of NR⁶, an optionallysubstituted C₁-C₆ alkyl, an optionally substituted C₃-C₈ cycloalkyl, anoptionally substituted C₁-C₆ heteroalkyl, and an optionally substitutednon-aromatic heterocycle;

L¹ is selected from NH and CHR²;

W is selected from O (oxygen) and NH;

X is N (nitrogen) or CR²;

Y is selected from an optionally substituted C₁-C₆ alkyl, C₂-C₆ alkenyl,an optionally substituted C₁-C₆ heteroalkyl, an optionally substitutedC₁-C₆ heteroalkenyl, an optionally substituted phenylalkenyl, and anoptionally substituted heterocyclealkenyl;

Z is O (oxygen) or S (sulfur);

n is 1, 2 or 3; and

provided that if R² is methyl, R⁴ is phenyl, L¹ is NH, and Q is N-Ph-R¹in Formula I and III, R¹ of Formula I and III is not selected from thegroup of halogen, alkyl, substituted alkyl, carboxylic acid, andcarboxylic esters.

In some embodiments, the compound is a GCSF receptor agonist.

In some embodiments, the compound is a GCSF receptor partial agonist.

Some embodiments include pharmaceutical compositions comprising acompound provided herein and a pharmaceutically acceptable excipient.

Some embodiments include methods of treating a hematopoietic orneurological disorder comprising administering an effective amount of acompound provided herein to a subject in need thereof.

In some embodiments, the disorder is selected from the group consistingof granulocytopenia, neutropenia, amyotrophic lateral sclerosis,multiple sclerosis, multiple dystrophy, and spinal cord injury.

In some embodiments, the compound is administered in combination with anadditional therapeutic regimen. In some embodiments, the additionaltherapeutic regimen is selected from the group consisting ofchemotherapy, bone marrow transplantation, and radiation therapy.

Some embodiments include methods of treating a hematopoietic orneurological disorder comprising administering an effective amount of acompound of Formula (V) or (VI) to a subject in need thereof, whereinFormula (V) or (VI) having the structure:

or pharmaceutically acceptable salt thereof, wherein:

R¹⁰ is selected from an optionally substituted C₁-C₁₂ alkyl, anoptionally substituted C₁-C₁₂ heteroalkyl, an optionally substitutedC₂-C₁₂ heteroalkenyl, an optionally substituted C₂-C₁₂ heteroalkynyl, anoptionally substituted C₁-C₁₂ cycloalkyl; an optionally substitutedC₁-C₆ heterocycloalkyl, an optionally substituted heteroarylalkyl, anoptionally substituted arylheteroalkyl, an optionally substitutedheteroarylheteroalkyl, OR¹⁴, SR¹⁴, and NR⁸R¹⁴;

R¹¹ is selected from hydrogen, halogen, an optionally substituted C₁-C₆alkyl, an optionally substituted C₁-C₆ heteroalkyl, an optionallysubstituted C₃-C₈ cycloalkyl, and an optionally substituted C₁-C₆heterocycle;

R¹² is selected from an optionally substituted C₁-C₆ heteroalkyl, anoptionally substituted C₃-C₈ cycloalkyl, an optionally substituted C₁-C₆heterocycle, an optionally substituted aryl, and an optionallysubstituted heteroaryl;

R¹³ is selected from hydrogen, halogen, an optionally substituted C₁-C₈alkyl, an optionally substituted C₁-C₈ heteroalkyl, an optionallysubstituted C₂-C₈ heteroalkenyl, an optionally substituted C₃-C₈cycloalkyl, an optionally substituted C₁-C₆ heterocycloalkyl, OR¹⁴,SR¹⁴, and NR⁸R¹⁴;

R¹⁴ is selected from an optionally substituted aryl and an optionallysubstituted heteroaryl;

V is selected from CH═CH, N═CH, CH═N, NH, O (oxygen), and S (sulfur);and

n is 1, 2 or 3.

In some such embodiments, the disorder is selected from the groupconsisting of granulocytopenia, neutropenia, amyotrophic lateralsclerosis, multiple sclerosis, multiple dystrophy, and spinal cordinjury.

In some embodiments, the compound is administered in combination with anadditional therapeutic regimen. In some embodiments, the additionaltherapeutic regimen is selected from the group consisting ofchemotherapy, bone marrow transplantation, and radiation therapy.

Some embodiments include methods for identifying a therapeutic compoundcomprising: contacting a target cell with a test compound comprising aGCSF receptor agonist, wherein the target cell comprises a mutant GCSFreceptor protein; and determining whether the test compoundsignificantly changes the level of activity of the mutant GCSF receptorin the target cell. Some such embodiments also include comparing thelevel of activity of the mutant GCSF receptor in the target cell to thelevel of activity of a wild type GCSF receptor protein in a cellcontacted with the test compound. Some such embodiments also includedetermining whether the level of activity of the mutant GCSF receptor inthe target cell contacted with the test compound is less than the levelof activity of the wild type GCSF receptor protein in a cell contactedwith the test compound.

In some embodiments, the mutant GCSF receptor protein comprises amutation in the transmembrane domain of the protein or a mutationproximal to the transmembrane domain of the protein. In some suchembodiments, the mutation is selected from the group consisting of asubstitution at the residue corresponding to his-627 of the human GCSFreceptor protein, and a substitution at the residue corresponding toAsp-602 of the mouse GCSF receptor protein.

In some embodiments, the mutant GCSF receptor comprises a mutant humanGCSF receptor.

In some embodiments, the cell comprises a mammalian cell. In someembodiments, the cell comprises a human cell. In some embodiments, thecell is not a blood cell.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A is a graph of relative luciferase activity in HepG2 cellstransfected with a STAT3-responsive reporter construct and treated withvarious concentrations of rhGCSF, Compound X, or control. FIG. 1B is agraph of luciferase activity in HEK293 cells transfected with aSTAT5-responsive reporter construct and treated with variousconcentrations of rhGCSF, Compound X, or control. Mean RLU (duplicates)±SD.

FIG. 2A and FIG. 2B are graphs of relative luciferase activity in HEK293cells transfected with a STAT5-responsive reporter construct and ahGCSFR, hTPOR, or hEPOR expression vector, and treated with rhGCSF,Compound X, or control. Mean (triplicates) ±SD.

FIG. 3A is a graph of relative luciferase activity of viability ofUTP-hGCSFR stable cells treated with rhGCSF or Compound X. FIG. 3B andFIG. 3C depict graphs of fold induction of STAT5 phosphorylation (upperpanel) or STAT3 phosphorylation (lower panel) in UTP-hGCSFR stable cellstreated with rhGCSF or Compound X. Mean (duplicates) ±SD.

FIG. 4 is a graph showing the relative expression of CD15 in CD34+ humanbone marrow cells treated with various concentrations of rhGCSF,Compound X, or rhGCSF and Compound X.

FIG. 5 is a schematic comparison of amino acid sequences of thetransmembrane domain of GCSFR from various species. Compound X is activein cells expressing human and monkey GCSFR, but not mouse, guinea pig,and rabbit receptors.

FIGS. 6A and 6B are graphs of relative STAT3-responsive luciferasereporter activity in cells treated with various concentrations ofrhGCSF, Compound X, or control, and transfected with various expressionvectors. FIG. 6A: left panel: hGCSFR; right panel: hGCSFR-H627N. FIG.6B: left panel: mGCSFR; right panel: mGCSFR-N607H.

FIG. 7 is a graph showing allosteric binding of Compound X to hGCSFR inUTP-hGCSFR cells. Compound X did not displace [¹²⁵I]rhGCSF and bindingof [¹²⁵I]rhGCSF was augmented in a concentration dependent manner.

DETAILED DESCRIPTION

Some embodiments include compositions and methods of using oridentifying compounds that modulate the activity of the granulocytecolony-stimulating factor receptor (GCFR). Some embodiments include useof compounds to treat certain disorders, such as hematopoietic orneurological disorders.

Certain embodiments include a compound of Formula (I), (II), (III), or(IV):

a tautomer thereof, or a pharmaceutically acceptable salt thereof,wherein:

R¹ is selected from hydrogen, OR⁶, NO₂, CN, NR⁶R⁷, CO₂R⁶, C(═O)NR⁶R⁷,SO₃R⁶, SO₂NR⁶R⁸, an optionally substituted C₁-C₆ alkyl, an optionallysubstituted C₂-C₆ alkenyl, an optionally substituted C₂-C₆ alkynyl, anoptionally substituted C₁-C₆ heteroalkyl, an optionally substitutedC₃-C₆ cycloalkyl, an optionally substituted C₃-C₆ cycloalkenyl, anoptionally substituted C₂-C₆ heterocyclyl, an optionally substitutedarylalkyl, an optionally substituted aryl, and an optionally substitutedheteroaryl;

R² and R³ are independently selected from hydrogen, an optionallysubstituted C₁-C₆ alkyl, an optionally substituted C₂-C₆ alkenyl, anoptionally substituted C₂-C₆ alkynyl, an optionally substituted C₁-C₆heteroalkyl, an optionally substituted C₃-C₈ cycloalkyl, an optionallysubstituted C₃-C₈ cycloalkenyl, an optionally substituted C₁-C₆heterocycle, an optionally substituted aryl, and an optionallysubstituted heteroaryl;

R⁴ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₂-C₆ alkenyl, an optionally substituted C₂-C₆alkynyl, an optionally substituted C₁-C₆ heteroalkyl, an optionallysubstituted C₃-C₈ cycloalkyl, an optionally substituted C₃-C₈cycloalkenyl, an optionally substituted C₁-C₆ heterocycle, an optionallysubstituted aryl, an optionally substituted heteroaryl, an optionallysubstituted arylalkyl, an optionally substituted arylalkenyl, anoptionally substituted arylalkynyl, and an optionally substitutedheteroarylalkyl;

R⁵ is selected from hydrogen, halogen, NO₂, CN, CF₃, OR⁶, CO₂R⁶,C(═O)NR⁶R⁷, SO₃R⁶, and SO₂NR⁶R⁸, an optionally substituted aryl, anoptionally substituted C₁-C₆ alkyl, and an optionally substituted C₁-C₆heteroalkyl;

R⁶ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, aC₁-C₆ heteroalkyl, an optionally substituted aryl, and an optionallysubstituted heteroaryl;

R⁷ is selected from hydrogen, C(═O)R⁸, C(═O)NHR⁸, an optionallysubstituted C₁-C₆ alkyl, and an optionally substituted C₁-C₆heteroalkyl; or —NR⁶R⁷ is an optionally substituted non-aromaticheterocycle linked through a ring nitrogen;

R⁸ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, andan optionally substituted C₁-C₆ heteroalkyl;

R⁹ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₂-C₆ alkenyl, an optionally substituted C₂-C₆alkynyl, an optionally substituted C₁-C₆ heteroalkyl, an optionallysubstituted C₃-C₈ cycloalkyl, an optionally substituted C₃-C₈cycloalkenyl, an optionally substituted C₁-C₆ heterocycle, an optionallysubstituted heteroaryl, an optionally substituted arylalkyl, anoptionally substituted arylalkenyl, an optionally substitutedarylalkynyl, an optionally substituted heteroarylalkyl, an optionallysubstituted heteroarylalkenyl, and an optionally substitutedheteroarylalkynyl;

Q is selected from the group consisting of NR⁶, an optionallysubstituted C₁-C₆ alkyl, an optionally substituted C₃-C₈ cycloalkyl, anoptionally substituted C₁-C₆ heteroalkyl, and an optionally substitutednon-aromatic heterocycle;

L¹ is selected from NH and CHR²;

W is selected from O (oxygen) and NH;

X is N (nitrogen) or CR²;

Y is selected from an optionally substituted C₁-C₆ alkyl, C₂-C₆ alkenyl,an optionally substituted C₁-C₆ heteroalkyl, an optionally substitutedC₁-C₆ heteroalkenyl, an optionally substituted phenylalkenyl, and anoptionally substituted heterocyclealkenyl;

Z is O (oxygen) or S (sulfur);

n is 1, 2 or 3; and

provided that if R² is methyl, R⁴ is phenyl, L¹ is NH, and Q is N-Ph-R¹in Formula I and III, R¹ of Formula I and III is not selected from thegroup of halogen, alkyl, substituted alkyl, carboxylic acid, andcarboxylic esters.

Certain embodiments include methods of treating a hematopoietic orneurological disorder comprising administering an effective amount of acompound of Formula (V) or (VI) to a subject in need thereof, whereinFormula (V) or (VI):having the structure:

or pharmaceutically acceptable salt thereof, wherein:

R¹⁰ is selected from an optionally substituted C₁-C₁₂ alkyl, anoptionally substituted C₁-C₁₂ heteroalkyl, an optionally substitutedC₂-C₁₂ heteroalkenyl, an optionally substituted C₂-C₁₂ heteroalkynyl, anoptionally substituted C₁-C₁₂ cycloalkyl; an optionally substitutedC₁-C₆ heterocycloalkyl, an optionally substituted heteroarylalkyl, anoptionally substituted arylheteroalkyl, an optionally substitutedheteroarylheteroalkyl, OR¹⁴, SR¹⁴, and NR⁸R¹⁴;

R¹¹ is selected from hydrogen, halogen, an optionally substituted C₁-C₆alkyl, an optionally substituted C₁-C₆ heteroalkyl, an optionallysubstituted C₃-C₈ cycloalkyl, and an optionally substituted C₁-C₆heterocycle;

R¹² is selected from an optionally substituted C₁-C₆ heteroalkyl, anoptionally substituted C₃-C₈ cycloalkyl, an optionally substituted C₁-C₆heterocycle, an optionally substituted aryl, and an optionallysubstituted heteroaryl;

R¹³ is selected from hydrogen, halogen, an optionally substituted C₁-C₈alkyl, an optionally substituted C₁-C₈ heteroalkyl, an optionallysubstituted C₂-C₈ heteroalkenyl, an optionally substituted C₃-C₈cycloalkyl, an optionally substituted C₁-C₆ heterocycloalkyl, OR¹⁴,SR¹⁴, and NR⁸R¹⁴;

R¹⁴ is selected from an optionally substituted aryl and an optionallysubstituted heteroaryl;

V is selected from CH═CH, N═CH, CH═N, NH, O (oxygen), and S (sulfur);and

n is 1, 2 or 3.

In certain embodiments, a compound of Formula I, II, III, IV, V, and VIis a GCSFR agonist. In some such embodiments, a compound of Formula I,II, III, IV, V, and VI is a hGCSFR agonist.

Compounds provided herein can have a variety of functions. For example,in certain embodiments, a compound of Formula I, II, III, IV, V, and VIis a GCSFR partial agonist. In some such embodiments, a compound ofFormula I, II, III, IV, V, and VI is a hGCSFR partial agonist. Incertain embodiments provide a selective GCSFR modulator. In certainembodiments provide a selective GCSF receptor agonist. In certainembodiments provide a selective GCSFR partial agonist. In certainembodiments provide a selective GCSF receptor binding compound. Incertain embodiments, selective GCSFR modulators are agonists, partialagonists, and/or antagonists for the GCSF receptor.

Definitions

Unless specific definitions are provided, the nomenclatures utilized inconnection with, and the laboratory procedures and techniques of,analytical chemistry, synthetic organic chemistry, and medicinal andpharmaceutical chemistry described herein are those known in the art.Standard chemical symbols are used interchangeably with the full namesrepresented by such symbols. Thus, for example, the terms “hydrogen” and“H” are understood to have identical meaning. Standard techniques may beused for chemical syntheses, chemical analyses, pharmaceuticalpreparation, formulation, and delivery, and treatment of patients.Standard techniques may be used for recombinant DNA, oligonucleotidesynthesis, and tissue culture and transformation (e.g., electroporation,lipofection). Reactions and purification techniques may be performede.g., using kits according to manufacturer's specifications or ascommonly accomplished in the art or as described herein. The foregoingtechniques and procedures may be generally performed according toconventional methods well known in the art and as described in variousgeneral and more specific references that are cited and discussedthroughout the present specification. See e.g., Sambrook et al.Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring HarborLaboratory Press, Cold Spring Harbor, N.Y. (1989)), which isincorporated herein by reference in its entirety for any purpose.

The term “selective binding compound” refers to a compound thatselectively binds to any portion of one or more target.

The term “binding-site selective hGCSFR activators” refers to a compoundthat selectively binds to a hGCSF receptor at or near TM domain.

The term “selectively binds” refers to the ability of a selectivebinding compound to bind to a target receptor with greater affinity thanit binds to a non-target receptor. In certain embodiments, selectivebinding refers to binding to a target with an affinity that is at least10, 50, 100, 250, 500, or 1000 times greater than the affinity for anon-target.

The term “target receptor” refers to a receptor or a portion of areceptor capable of being bound by a selective binding compound. Incertain embodiments, a target receptor is a hGCSF receptor.

The term “modulator” refers to a compound that alters an activity. Forexample, a modulator may cause an increase or decrease in the magnitudeof a certain activity compared to the magnitude of the activity in theabsence of the modulator. In certain embodiments, a modulator is aninhibitor, which decreases the magnitude of one or more activities. Incertain embodiments, an inhibitor completely prevents one or morebiological activities. In certain embodiments, a modulator is anactivator, which increases the magnitude of at least one activity. Incertain embodiments the presence of a modulator results in a activitythat does not occur in the absence of the modulator.

The term “selective modulator” refers to a compound that selectivelymodulates a target activity.

The term “selective hGCSFR modulator” refers to a compound thatselectively modulates hGCSFR activity. The term selective hGCSFRmodulator includes, but is not limited to “hGCSF mimic” which refers toa compound, the presence of which results in similar GCSF activity.

The term “selectively modulates” refers to the ability of a selectivemodulator to modulate a target activity to a greater extent than itmodulates a non-target activity.

The term “target activity” refers to a biological activity capable ofbeing modulated by a selective modulator. Certain exemplary targetactivities include, but are not limited to, binding affinity, signaltransduction, enzymatic activity, the proliferation and/ordifferentiation of progenitor cells, generation of white blood cells,and alleviation of symptoms of a disease or condition.

The term “GCSF activity” refers to a biological activity that results,either directly or indirectly from the presence of GCSF. Example GCSFactivities include, but are not limited to, proliferation and/ordifferentiation of progenitor cells to produce white blood cells;hematopoiesis; growth and/or development of glial cells; repair of nervecells; and alleviation of granulocytopenia.

The term “receptor mediated activity” refers to any biological activitythat results, either directly or indirectly, from binding of a ligand toa receptor.

The term “agonist” refers to a compound, the presence of which resultsin a biological activity of a receptor that is the same as thebiological activity resulting from the presence of a naturally occurringligand for the receptor.

The term “partial agonist” refers to a compound, the presence of whichresults in a biological activity of a receptor that is of the same typeas that resulting from the presence of a naturally occurring ligand forthe receptor, but of a lower magnitude.

The term “antagonist” refers to a compound, the presence of whichresults in a decrease in the magnitude of a biological activity of areceptor. In certain embodiments, the presence of an antagonist resultsin complete inhibition of a biological activity of a receptor.

The term “alkyl” refers to a branched or unbranched fully saturatedacyclic aliphatic hydrocarbon group. An alkyl may be branched orstraight chain. Alkyls may be substituted or unsubstituted. Alkylsinclude, but are not limited to, methyl, ethyl, propyl, isopropyl,butyl, isobutyl, tertiary butyl, pentyl, hexyl, and the like, each ofwhich may be optionally substituted.

In certain embodiments, an alkyl comprises 1 to 20 carbon atoms(whenever it appears herein, a numerical range such as “1 to 20” refersto each integer in the given range; e.g., “1 to 20 carbon atoms” meansthat an alkyl group may comprise only 1 carbon atom, 2 carbon atoms, 3carbon atoms, etc., up to and including 20 carbon atoms, although theterm “alkyl” also includes instances where no numerical range of carbonatoms is designated). An alkyl may be designated as “C₁-C₆ alkyl” orsimilar designations. By way of example only, “C₁-C₄ alkyl” indicates analkyl having one, two, three, or four carbon atoms, e.g., the alkyl isselected from methyl, ethyl, propyl, iso-propyl, butyl, iso-butyl,sec-butyl, and tent-butyl.

The term “alkenyl” used herein refers to a monovalent straight orbranched chain aliphatic hydrocarbon radical of from two to twentycarbon atoms containing at least one carbon-carbon double bondincluding, but not limited to, 1-propenyl, 2-propenyl,2-methyl-1-propenyl, 1-butenyl, 2-butenyl, and the like. In certainembodiments, an alkenyl comprises 2 to 20 carbon atoms (whenever itappears herein, a numerical range such as “2 to 20” refers to eachinteger in the given range; e.g., “2 to 20 carbon atoms” means that analkenyl group may comprise only 2 carbon atoms, 3 carbon atoms, etc., upto and including 20 carbon atoms, although the term “alkenyl” alsoincludes instances where no numerical range of carbon atoms isdesignated). An alkenyl may be designated as “C₂-C₆ alkenyl” or similardesignations. By way of example only, “C₂-C₄ alkenyl” indicates analkenyl having two, three, or four carbon atoms, e.g., the alkenyl isselected from ethenyl, propenyl, and butenyl.

The term “alkynyl” used herein refers to a monovalent straight orbranched chain aliphatic hydrocarbon radical of from two to twentycarbon atoms containing at least one carbon-carbon triple bondincluding, but not limited to, 1-propynyl, 1-butynyl, 2-butynyl, and thelike. In certain embodiments, an alkynyl comprises 2 to 20 carbon atoms(whenever it appears herein, a numerical range such as “2 to 20” refersto each integer in the given range; e.g., “2 to 20 carbon atoms” meansthat an alkynyl group may comprise only 2 carbon atoms, 3 carbon atoms,etc., up to and including 20 carbon atoms, although the term “alkynyl”also includes instances where no numerical range of carbon atoms isdesignated). An alkynyl may be designated as ^(“)C₂-C₆ alkynyl” orsimilar designations. By way of example only, “C₂-C₄ alkynyl” indicatesan alkenyl having two, three, or four carbon atoms, e.g., the alkenyl isselected from ethynyl, propynyl, and butynyl.

The term “cycloalkyl” used herein refers to saturated aliphatic ringsystem radical having three to twenty carbon atoms. A cycloalkyl refersto monocyclic and polycyclic saturated aliphatic ring system including,but not limited to, cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl,bicyclo[4.4.0]decanyl, bicyclo[2.2.1]heptanyl, adamantyl, norbornyl, andthe like. In certain embodiments, a cycloalkyl comprises 3 to 20 carbonatoms (whenever it appears herein, a numerical range such as “3 to 20”refers to each integer in the given range; e.g., “3 to 20 carbon atoms”means that a cycloalkyl group may comprise only 3 carbon atoms, etc., upto and including 20 carbon atoms, although the term “cycloalkyl” alsoincludes instances where no numerical range of carbon atoms isdesignated). A cycloalkyl may be designated as “C₃-C₇ cycloalkyl” orsimilar designations. By way of example only, “C₃-C₆ cycloalkyl”indicates an alkenyl having two, three, four, five or six carbon atoms,e.g., the cycloalkyl is selected from cyclopropyl, cyclobutyl,cyclopentyl, and cyclohexyl.

The term “cycloalkenyl” used herein refers to aliphatic ring systemradical having three to twenty carbon atoms having at least onecarbon-carbon double bond in the ring. A cycloalkenyl refers tomonocyclic and polycyclic unsaturated aliphatic ring system including,but are not limited to, cyclopropenyl, cyclopentenyl, cyclohexenyl,cycloheptenyl, bicyclo[3.1.0]hexyl, norbornylenyl, 1,1′-bicyclopentenyl,and the like. In certain embodiments, a cycloalkenyl comprises 3 to 20carbon atoms (whenever it appears herein, a numerical range such as “3to 20” refers to each integer in the given range; e.g., “3 to 20 carbonatoms” means that a cycloalkenyl group may comprise only 3 carbon atoms,etc., up to and including 20 carbon atoms, although the term“cycloalkenyl” also includes instances where no numerical range ofcarbon atoms is designated). A cycloalkenyl may be designated as “C₃-C₇cycloalkenyl” or similar designations. By way of example only, “C₃-C₆cycloalkenyl” indicates an alkenyl having two, three, four, five or sixcarbon atoms, e.g., the cycloalkyl is selected from cyclopropenyl,cyclobutenyl, cyclopentenyl, and cyclohexenyl.

The term “heteroalkyl” refers to a group comprising an alkyl and one ormore heteroatoms. Certain heteroalkyls are acylalkyls, in which the oneor more heteroatoms are within an alkyl chain. Examples of heteroalkylsinclude, but are not limited to, CH₃C(═O)CH₂—, CH₃C(═O)CH₂CH₂—,CH₃CH₂C(═O)CH₂CH₂—, CH₃C(═O)CH₂CH₂CH₂—, CH₃OCH₂CH₂—, CH₃NHCH₂—, and thelike.

The term “alkoxy” used herein refers to straight or branched chain alkylradical covalently bonded to the parent molecule through an —O— linkage.Examples of alkoxy groups include, but are not limited to, methoxy,ethoxy, propoxy, isopropoxy, butoxy, n-butoxy, sec-butoxy, t-butoxy andthe like. An alkoxy may be designated as “C₁-C₆ alkoxy” or similardesignations. By way of example only, “C₁-C₄ alkoxy” indicates an alkylhaving one, two, three, or four carbon atoms, e.g., the alkoxy isselected from methoxy, ethoxy, propoxy, iso-propoxy, butoxy, iso-butoxy,sec-butoxy, and tent-butoxy.

The term “olefin” refers to a C═C bond.

The term “alkylideneamino” used herein refers to a moiety of from one totwenty carbon atoms containing at least one carbon-nitrogen double bondwhere the moiety is connected to the main group through the nitrogen,including, but not limited to, methylideneamino, ethylideneamino,methylethylideneamino, propylideneamino, 1-methylpropylideneaminyl,2-methylpropylideneamino, butylideneamino, 1-methylbutylideneamino,2-methylbutylideneamino, cyclopropylideneamino, cyclobutylideneamino,cyclopentylideneamino, cyclohexylideneamino and the like.

The term “carbocycle” refers to a group comprising a covalently closedring, wherein each of the atoms forming the ring is a carbon atom.Carbocyclic rings may be formed by three, four, five, six, seven, eight,nine, or more than nine carbon atoms. Carbocycles may be optionallysubstituted.

The term “heterocycle” refers to a group comprising a covalently closedring wherein at least one atom forming the ring is a heteroatom.Heterocyclic rings may be formed by three, four, five, six, seven,eight, nine, or more than nine atoms. Any number of those atoms may beheteroatoms (i.e., a heterocyclic ring may comprise one, two, three,four, five, six, seven, eight, nine, or more than nine heteroatoms). Inheterocyclic rings comprising two or more heteroatoms, those two or moreheteroatoms may be the same or different from one another. Heterocyclesmay be optionally substituted. Binding to a heterocycle can be at aheteroatom or via a carbon atom. For example, binding for benzo-fusedderivatives, may be via a carbon of the benzenoid ring. Examples ofheterocycles include, but are not limited to the following:

wherein D, E, F, and G independently represent a heteroatom. Each of D,E, F, and G may be the same or different from one another.

The term “heteroatom” refers to an atom other than carbon or hydrogen.Heteroatoms are typically independently selected from oxygen, sulfur,nitrogen, and phosphorus, but are not limited to those atoms. Inembodiments in which two or more heteroatoms are present, the two ormore heteroatoms may all be the same as one another, or some or all ofthe two or more heteroatoms may each be different from the others.

The term “aromatic” refers to a group comprising a covalently closedring having a delocalized π-electron system. Aromatic rings may beformed by five, six, seven, eight, nine, or more than nine atoms.Aromatics may be optionally substituted. Examples of aromatic groupsinclude, but are not limited to phenyl, naphthalenyl, phenanthrenyl,anthracenyl, tetralinyl, fluorenyl, indenyl, and indanyl. The termaromatic includes, for example, benzenoid groups, connected via one ofthe ring-forming carbon atoms, and optionally carrying one or moresubstituents selected from an aryl, a heteroaryl, a cycloalkyl, anon-aromatic heterocycle, a halo, a hydroxy, an amino, a cyano, a nitro,an alkylamido, an acyl, a C₁₋₆ alkoxy, a C₁₋₆ alkyl, a C₁₋₆hydroxyalkyl, a C₁₋₆ aminoalkyl, a C₁₋₆ alkylamino, an alkylsulfenyl, analkylsulfinyl, an alkylsulfonyl, an sulfamoyl, or a trifluoromethyl. Incertain embodiments, an aromatic group is substituted at one or more ofthe para, meta, and/or ortho positions. Examples of aromatic groupscomprising substitutions include, but are not limited to, phenyl,3-halophenyl, 4-halophenyl, 3-hydroxyphenyl, 4-hydroxyphenyl,3-aminophenyl, 4-aminophenyl, 3-methylphenyl, 4-methylphenyl,3-methoxyphenyl, 4-methoxyphenyl, 4-trifluoromethoxyphenyl,3-cyanophenyl, 4-cyanophenyl, dimethylphenyl, naphthyl, hydroxynaphthyl,hydroxymethylphenyl, (trifluoromethyl)phenyl, alkoxyphenyl,4-morpholin-4-ylphenyl, 4-pyrrolidin-1-ylphenyl, 4-pyrazolylphenyl,4-triazolylphenyl, and 4-(2-oxopyrrolidin-1-yl)phenyl.

The term “aryl” refers to an aromatic group wherein each of the atomsforming the ring is a carbon atom. Aryl rings may be formed by five,six, seven, eight, nine, or more than nine carbon atoms. Aryl groups maybe optionally substituted.

The term “heteroaryl” refers to an aromatic group wherein at least oneatom forming the aromatic ring is a heteroatom. Heteroaryl rings may beformed by three, four, five, six, seven, eight, nine, or more than nineatoms. Heteroaryl groups may be optionally substituted. Examples ofheteroaryl groups include, but are not limited to, aromatic C₃₋₈heterocyclic groups comprising one oxygen or sulfur atom or up to fournitrogen atoms, or a combination of one oxygen or sulfur atom and up totwo nitrogen atoms, and their substituted as well as benzo- andpyrido-fused derivatives, for example, connected via one of thering-forming carbon atoms. In certain embodiments, heteroaryl groups areoptionally substituted with one or more substituents, independentlyselected from halo, hydroxy, amino, cyano, nitro, alkylamido, acyl,C₁₋₆-alkoxy, C₁₋₆-alkyl, C₁₋₆-hydroxyalkyl, C₁₋₆-aminoalkyl,C₁₋₆-alkylamino, alkylsulfenyl, alkylsulfinyl, alkylsulfonyl, sulfamoyl,or trifluoromethyl. Examples of heteroaryl groups include, but are notlimited to, unsubstituted and mono- or di-substituted derivatives offuran, benzofuran, thiophene, benzothiophene, pyrrole, pyridine, indole,oxazole, benzoxazole, isoxazole, benzisoxazole, thiazole, benzothiazole,isothiazole, imidazole, benzimidazole, pyrazole, indazole, tetrazole,quinoline, isoquinoline, pyridazine, pyrimidine, purine and pyrazine,furazan, 1,2,3-oxadiazole, 1,2,3-thiadiazole, 1,2,4-thiadiazole,triazole, benzotriazole, pteridine, phenoxazole, oxadiazole,benzopyrazole, quinolizine, cinnoline, phthalazine, quinazoline, andquinoxaline. In some embodiments, the substituents are halo, hydroxy,cyano, O—C₁₋₆-alkyl, C₁₋₆-alkyl, hydroxy-C₁₋₆-alkyl, andamino-C₁₋₆-alkyl.

The term “non-aromatic ring” refers to a group comprising a covalentlyclosed ring that does not have a delocalized π-electron system.

The term “non-aromatic heterocycle” refers to a group comprising anon-aromatic ring wherein one or more atoms forming the ring is aheteroatom. Non-aromatic heterocyclic rings may be formed by three,four, five, six, seven, eight, nine, or more than nine atoms.Non-aromatic heterocycles may be optionally substituted. In certainembodiments, non-aromatic heterocycles comprise one or more carbonyl orthiocarbonyl groups such as, for example, oxo- and thio-containinggroups. Examples of non-aromatic heterocycles include, but are notlimited to, lactams, lactones, cyclic imides, cyclic thioimides, cycliccarbamates, tetrahydrothiopyran, 4H-pyran, tetrahydropyran, piperidine,1,3-dioxin, 1,3-dioxane, 1,4-dioxin, 1,4-dioxane, piperazine,1,3-oxathiane, 1,4-oxathiin, 1,4-oxathiane, tetrahydro-1,4-thiazine,2H-1,2-oxazine, maleimide, succinimide, barbituric acid, thiobarbituricacid, dioxopiperazine, hydantoin, dihydrouracil, morpholine, trioxane,hexahydro-1,3,5-triazine, tetrahydrothiophene, tetrahydrofuran,pyrroline, pyrrolidine, pyrrolidone, pyrrolidinone, pyrazoline,pyrazolidine, imidazoline, imidazolidine, 1,3-dioxole, 1,3-dioxolane,1,3-dithiole, 1,3-dithiolane, isoxazoline, isoxazolidine, oxazoline,oxazolidine, oxazolidinone, thiazoline, thiazolidine, and1,3-oxathiolane.

The term “arylalkyl” refers to a group comprising an aryl group bound toan alkyl group.

The term “arylalkenyl” refers to a group comprising an aryl group boundto an alkenyl group.

The term “arylalkynyl” refers to a group comprising an aryl group boundto an alkynyl group.

The term “heteroarylalkyl” refers to a group comprising a heteroarylgroup bound to an alkyl group.

The term “heteroarylalkenyl” refers to a group comprising a heteroarylgroup bound to an alkenyl group.

The term “heteroarylalkynyl” refers to a group comprising a heteroarylgroup bound to an alkynyl group.

The term “carbocycloalkyl” refers to a group comprising a carbocycliccycloalkyl ring. Carbocycloalkyl rings may be formed by three, four,five, six, seven, eight, nine, or more than nine carbon atoms.Carbocycloalkyl groups may be optionally substituted.

The term “ring” refers to any covalently closed structure. Ringsinclude, for example, carbocycles (e.g., aryls and cycloalkyls),heterocycles (e.g., heteroaryls and non-aromatic heterocycles),aromatics (e.g., aryls and heteroaryls), and non-aromatics (e.g.,cycloalkyls and non-aromatic heterocycles). Rings may be optionallysubstituted. Rings may form part of a ring system.

The term “ring system” refers to a either a single ring or two or morerings, wherein, if two or more rings are present, the two or more of therings are fused. The term “fused” refers to structures in which two ormore rings share one or more bonds.

As used herein, the term “linked to form a ring” refers to instanceswhere two atoms that are bound either to a single atom or to atoms thatare themselves ultimately bound, are each bound to a linking group, suchthat the resulting structure forms a ring. That resulting ring comprisesthe two atoms that are linked to form a ring, the atom (or atoms) thatpreviously linked those atoms, and the linker. For example, if A and Ebelow are “linked to form a ring”

the resulting ring includes A, E, the C (carbon) or N (nitrogen) towhich they are attached, and a linking group. Unless otherwiseindicated, that linking group may be of any length and may be optionallysubstituted. Referring to the above example, resulting structuresinclude, but are not limited to:

and the like.

In certain embodiments, the two substituents that together form a ringare not immediately bound to the same atom. For example, if A and E,below, are linked to form a ring:

the resulting ring comprises A, E, the two atoms that already link A andE and a linking group. Examples of resulting structures include, but arenot limited to:

and the like.

In certain embodiments, the atoms that together form a ring areseparated by three or more atoms. For example, if A and E, below, arelinked to form a ring:

the resulting ring comprises A, E, the 3 atoms that already link A andE, and a linking group. Examples of resulting structures include, butare not limited to:

and the like.

As used herein, the term “together form a bond” refers to the instancein which two substituents to neighboring atoms are null the bond betweenthe neighboring atoms becomes a double bond. For example, if A and Ebelow “together form a bond”

the resulting structure is:

The term “null” refers to a group being absent from a structure. Forexample, in the structure

where in certain instances X is N (nitrogen), if X is N (nitrogen), oneof R′ or R″ is null, meaning that only three groups are bound to the N(nitrogen).

The substituent “R” appearing by itself and without a number designationrefers to a substituent selected from alkyl, cycloalkyl, aryl,heteroaryl (bonded through a ring carbon) and non-aromatic heterocycle(bonded through a ring carbon).

The term “O-carboxy” refers to the group consisting of formula RC(═O)O—.

The term “C-carboxy” refers to the group consisting of formula —C(═O)OR.

The term “acetyl” refers to the group consisting of formula —C(═O)CH₃.

The term “trihalomethanesulfonyl” refers to the group consisting offormula X₃CS(═O)₂— where X is a halogen.

The term “cyano” refers to the group consisting of formula —CN.

The term “isocyanato” refers to the group consisting of formula —NCO.

The term “thiocyanato” refers to the group consisting of formula —CNS.

The term “isothiocyanato” refers to the group consisting of formula—NCS.

The term “sulfonyl” refers to the group consisting of formula —S(═O)—R.

The term “S-sulfonamido” refers to the group consisting of formula—S(═O)₂NR.

The term “N-sulfonamido” refers to the group consisting of formulaRS(═O)₂NH—.

The term “trihalomethanesulfonamido” refers to the group consisting offormula X₃CS(═O)₂NR—.

The term “O-carbamyl” refers to the group consisting of formula—OC(═O)—NR.

The term “N-carbamyl” refers to the group consisting of formulaROC(═O)NH—.

The term “O-thiocarbamyl” refers to the group consisting of formula—OC(═S)—NR.

The term “N-thiocarbamyl” refers to the group consisting of formulaROC(═S)NH—.

The term “C-amido” refers to the group consisting of formula —C(═O)—NR₂.

The term “N-amido” refers to the group consisting of formula RC(═O)NH—.

The term “oxo” refers to the group consisting of formula ═O.

The term “keto” and “carbonyl” used herein refers to C═O.

The term “thiocarbonyl” used herein refers to C═S.

The term “ester” refers to a chemical moiety with formula—(R)_(n)—C(═O)OR′, where R and R′ are independently selected from alkyl,cycloalkyl, aryl, heteroaryl (bonded through a ring carbon) andnon-aromatic heterocycle (bonded through a ring carbon), where n is 0 or1.

The term “amide” refers to a chemical moiety with formula—(R)_(n)—C(═O)NHR′ or —(R)_(n)—NHC(═O)R′, where R is selected fromalkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon) andheteroalicyclic (bonded through a ring carbon), where n is 0 or 1 and R′is selected from hydrogen, alkyl, cycloalkyl, aryl, heteroaryl (bondedthrough a ring carbon) and heteroalicyclic (bonded through a ringcarbon), where n is 0 or 1. In certain embodiments, an amide may be anamino acid or a peptide.

The term “amino” refers to a chemical moiety with formula —NHR′R″, whereR′ and R″ are each independently selected from hydrogen, alkyl,cycloalkyl, aryl, heteroaryl (bonded through a ring carbon) andheteroalicyclic (bonded through a ring carbon).

The terms “amine,” “hydroxy,” and “carboxyl” include such groups thathave been esterified or amidified. Procedures and specific groups usedto achieve esterification and amidification are known to those of skillin the art and can readily be found in reference sources such as Greeneand Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley &Sons, New York, NY, 1999, which is incorporated herein in its entirety.

Unless otherwise indicated, the term “optionally substituted,” refers toa group in which none, one, or more than one of the hydrogen atoms hasbeen replaced with one or more group(s) individually and independentlyselected from: alkyl, alkenyl, cycloalkenyl, alkynyl, heteroalkyl,haloalkyl, cycloalkyl, aryl, arylalkyl, alkenylO—, arylalkylO—,arylalkylNH—, alkenylO—, cycloalkylC(═O)—, arylC(═O)—, arylC(═O)NH—,arylNHC(═O)—, aryl(CH₂)₀₋₃O(CH₂)₀₋₃—, —COOH, HO(CH₂)₁₋₃NH—,HO(CH₂)₁₋₃O—, HO(CH₂)₁₋₃—, HO(CH₂)₁₋₃O(CH₂)₁₋₃—, —C(═O)NHNH₂,heteroaryl, non-aromatic heterocycle, hydroxy, alkoxy, aryloxy,mercapto, alkylthio, arylthio, cyano, halo, carbonyl, oxo, thiocarbonyl,O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido,N-amido, S-sulfonamido, N-sulfonamido, C-carboxy, O-carboxy, isocyanato,thiocyanato, isothiocyanato, nitro, silyl, trihalomethanesulfonyl, andamino, including mono- and di-substituted amino groups, and theprotected derivatives of amino groups. Such protective derivatives (andprotecting groups that may form such protective derivatives) are knownto those of skill in the art and may be found in references such asGreene and Wuts, above. When the group contains a nitrogen, or a sulfur,an oxo as a substituent also includes oxides, for examplepyridine-N-oxide, thiopyran sulfoxide and thiopyran-S,S-dioxide. Inembodiments in which two or more hydrogen atoms have been substituted,the substituent groups may together form a ring.

The term “isomer” includes but not limited to stereoic isomers,geometric isomers, enantiomeric isomers, tautomeric isomers, andatromeric isomers.

Throughout the specification, groups and substituents thereof can bechosen by one skilled in the field to provide stable moieties andcompounds.

The term “carrier” refers to a compound that facilitates theincorporation of another compound into cells or tissues. For example,dimethyl sulfoxide (DMSO) is a commonly used carrier for improvingincorporation of certain organic compounds into cells or tissues.

The term “pharmaceutical agent” refers to a chemical compound orcomposition capable of inducing a desired therapeutic effect in apatient. In certain embodiments, a pharmaceutical agent comprises anactive agent, which is the agent that induces the desired therapeuticeffect. In certain embodiments, a pharmaceutical agent comprises aprodrug. In certain embodiments, a pharmaceutical agent comprisesinactive ingredients such as carriers, excipients, and the like.

The term “therapeutically effective amount” refers to an amount of apharmaceutical agent sufficient to achieve a desired therapeutic effect.

The term “prodrug” refers to a pharmaceutical agent that is convertedfrom a less active form into a corresponding more active form in vivo.

The term “pharmaceutically acceptable” refers to a formulation of acompound that does not significantly abrogate the biological activity, apharmacological activity and/or other properties of the compound whenthe formulated compound is administered to a patient. In certainembodiments, a pharmaceutically acceptable formulation does not causesignificant irritation to a patient.

The term “co-administer” refers to administering more than onepharmaceutical agent to a patient. In certain embodiments,co-administered pharmaceutical agents are administered together in asingle dosage unit. In certain embodiments, co-administeredpharmaceutical agents are administered separately. In certainembodiments, co-administered pharmaceutical agents are administered atthe same time. In certain embodiments, co-administered pharmaceuticalagents are administered at different times.

The term “patient” includes human and animal subjects.

The term “substantially pure” means an object species (e.g., compound)is the predominant species present (i.e., on a molar basis it is moreabundant than any other individual species in the composition). Incertain embodiments, a substantially purified fraction is a compositionwherein the object species comprises at least about 50 percent (on amolar basis) of all species present. In certain embodiments, asubstantially pure composition will comprise more than about 80%, 85%,90%, 95%, or 99% of all species present in the composition. In certainembodiments, the object species is purified to essential homogeneity(contaminant species cannot be detected in the composition byconventional detection methods) wherein the composition consistsessentially of a single species.

The term “tissue-selective” refers to the ability of a compound tomodulate a biological activity in one tissue to a greater or lesserdegree than it modulates a biological activity in another tissue. Thebiological activities in the different tissues may be the same or theymay be different. The biological activities in the different tissues maybe mediated by the same type of target receptor. For example, in certainembodiments, a tissue-selective compound may modulate receptor mediatedbiological activity in one tissue and fail to modulate, or modulate to alesser degree, receptor mediated biological activity in another tissuetype.

The term “monitoring” refers to observing an effect or absence of anyeffect. In certain embodiments, one monitors cells after contactingthose cells with a compound of the present embodiments. Examples ofeffects that may be monitored include, but are not limited to, changesin cell phenotype, cell proliferation, receptor activity, or theinteraction between a receptor and a compound known to bind to thereceptor.

The term “cell phenotype” refers to physical or biologicalcharacteristics. Examples of characteristics that constitute phenotypeincluded, but are not limited to, cell size, cell proliferation, celldifferentiation, cell survival, apoptosis (cell death), or theutilization of a metabolic nutrient (e.g., glucose uptake). Certainchanges or the absence of changes in cell phenotype are readilymonitored using techniques known in the art.

The term “cell proliferation” refers to the rate at which cells divide.In certain embodiments, cells are in situ in an organism. In certainembodiments, cell are grown in vitro in a vessel. The number of cellsgrowing in a vessel can be quantified by a person skilled in the art(e.g., by counting cells in a defined area using a microscope or byusing laboratory apparatus that measure the density of cells in anappropriate medium). One skilled in that art can calculate cellproliferation by determining the number of cells at two or more times.

The term “contacting” refers to bringing two or more materials intoclose enough proximity that they may interact. In certain embodiments,contacting can be accomplished in a vessel such as a test tube, a petridish, or the like. In certain embodiments, contacting may be performedin the presence of additional materials. In certain embodiments,contacting may be performed in the presence of cells. In certain of suchembodiments, one or more of the materials that are being contacted maybe inside a cell. Cells may be alive or may dead. Cells may or may notbe intact.

In certain embodiments, a salt corresponding to any of the compoundsprovided herein is provided. In certain embodiments, a saltcorresponding to a selective GCSFR modulator is provided. In certainembodiments, a salt corresponding to a selective GCSF receptor bindingagent is provided. In certain embodiments, a salt is obtained byreacting a compound with an acid, such as hydrochloric acid, hydrobromicacid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid,ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and thelike. In certain embodiments, a salt is obtained by reacting a compoundwith a base to form a salt such as an ammonium salt, an alkali metalsalt, such as a sodium or a potassium salt, an alkaline earth metalsalt, such as a calcium or a magnesium salt, a salt of organic basessuch as choline, dicyclohexylamine, N-methyl-D-glucamine,tris(hydroxymethyl)methylamine, 4-(2-hydroxyethyl)-morpholine,1-(2-hydroxyethyl)-pyrrolidine, ethanolamine and salts with amino acidssuch as arginine, lysine, and the like. In certain embodiments, a saltis obtained by reacting a free acid form of a selective HGF modulator orselective GCSFR binding agent with multiple molar equivalents of a base,such as bis-sodium, bis-ethanolamine, and the like.

In certain embodiments, a salt corresponding to a compound of thepresent embodiments is selected from acetate, ammonium,benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate,bromide, calcium edetate, camsylate, carbonate, chloride, cholinate,clavulanate, citrate, dihydrochloride, diphosphate, edetate, edisylate,estolate, esylate, fumarate, gluceptate, gluconate, glutamate,glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide,hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate,lactobionate, laurate, magnesium, malate, maleate, mandelate, mucate,napsylate, nitrate, N-methylglucamine, oxalate, pamoate (embonate),palmitate, pantothenate, phosphate, polygalacturonate, potassium,salicylate, sodium, stearate, subacetate, succinate, sulfate, tannate,tartrate, teoclate, tosylate, triethiodide, tromethamine,trimethylammonium, and valerate salts.

In certain embodiments, one or more carbon atoms of a compound of thepresent embodiments are replaced with silicon. See e.g., WO 03/037905A1;Tacke and Zilch, Endeavour, New Series, 10, 191-197 (1986); Bains andTacke, Curr. Opin. Drug Discov Devel. Jul:6(4):526-43(2003), all ofwhich are incorporated herein by reference in their entirety. In certainembodiments, compounds comprising one or more silicon atoms possesscertain desired properties, including, but not limited to, greaterstability and/or longer half-life in a patient, when compared to thesame compound in which none of the carbon atoms have been replaced witha silicon atom.

Certain compounds

Certain embodiments include a compound of Formula (I), (II), (III), or(IV):

a tautomer thereof, or a pharmaceutically acceptable salt thereof,wherein:

R¹ is selected from hydrogen, OR⁶, NO₂, CN, NR⁶R⁷, CO₂R⁶, C(═O)NR⁶R⁷,SO₃R⁶, SO₂NR⁶R⁸, an optionally substituted C₁-C₆ alkyl, an optionallysubstituted C₂-C₆ alkenyl, an optionally substituted C₂-C₆ alkynyl, anoptionally substituted C₁-C₆ heteroalkyl, an optionally substitutedC₃-C₆ cycloalkyl, an optionally substituted C₃-C₆ cycloalkenyl, anoptionally substituted C₂-C₆ heterocyclyl, an optionally substitutedarylalkyl, an optionally substituted aryl, and an optionally substitutedheteroaryl;

R² and R³ are independently selected from hydrogen, an optionallysubstituted C₁-C₆ alkyl, an optionally substituted C₂-C₆ alkenyl, anoptionally substituted C₂-C₆ alkynyl, an optionally substituted C₁-C₆heteroalkyl, an optionally substituted C₃-C₈ cycloalkyl, an optionallysubstituted C₃-C₈ cycloalkenyl, an optionally substituted C₁-C₆heterocycle, an optionally substituted aryl, and an optionallysubstituted heteroaryl;

R⁴ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₂-C₆ alkenyl, an optionally substituted C₂-C₆alkynyl, an optionally substituted C₁-C₆ heteroalkyl, an optionallysubstituted C₃-C₈ cycloalkyl, an optionally substituted C₃-C₈cycloalkenyl, an optionally substituted C₁-C₆ heterocycle, an optionallysubstituted aryl, an optionally substituted heteroaryl, an optionallysubstituted arylalkyl, an optionally substituted arylalkenyl, anoptionally substituted arylalkynyl, and an optionally substitutedheteroarylalkyl;

R⁵ is selected from hydrogen, halogen, NO₂, CN, CF₃, OR⁶, CO₂R⁶,C(═O)NR⁶R⁷, SO₃R⁶, and SO₂NR⁶R⁸, an optionally substituted aryl, anoptionally substituted C₁-C₆ alkyl, and an optionally substituted C₁-C₆heteroalkyl;

R⁶ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, aC₁-C₆ heteroalkyl, an optionally substituted aryl, and an optionallysubstituted heteroaryl;

R⁷ is selected from hydrogen, C(═O)R⁸, C(═O)NHR⁸, an optionallysubstituted C₁-C₆ alkyl, and an optionally substituted C₁-C₆heteroalkyl; or —NR⁶R⁷ is an optionally substituted non-aromaticheterocycle linked through a ring nitrogen;

R⁸ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, andan optionally substituted C₁-C₆ heteroalkyl;

R⁹ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₂-C₆ alkenyl, an optionally substituted C₂-C₆alkynyl, an optionally substituted C₁-C₆ heteroalkyl, an optionallysubstituted C₃-C₈ cycloalkyl, an optionally substituted C₃-C₈cycloalkenyl, an optionally substituted C₁-C₆ heterocycle, an optionallysubstituted heteroaryl, an optionally substituted arylalkyl, anoptionally substituted arylalkenyl, an optionally substitutedarylalkynyl, an optionally substituted heteroarylalkyl, an optionallysubstituted heteroarylalkenyl, and an optionally substitutedheteroarylalkynyl;

Q is selected from the group consisting of NR⁶, an optionallysubstituted C₁-C₆ alkyl, an optionally substituted C₃-C₈ cycloalkyl, anoptionally substituted C₁-C₆ heteroalkyl, and an optionally substitutednon-aromatic heterocycle;

L¹ is selected from NH and CHR²;

W is selected from O (oxygen) and NH;

X is N (nitrogen) or CR²;

Y is selected from an optionally substituted C₁-C₆ alkyl, C₂-C₆ alkenyl,an optionally substituted C₁-C₆ heteroalkyl, an optionally substitutedC₁-C₆ heteroalkenyl, an optionally substituted phenylalkenyl, and anoptionally substituted heterocyclealkenyl;

Z is O (oxygen) or S (sulfur);

n is 1, 2 or 3; and

provided that if R² is methyl, R⁴ is phenyl, L¹ is NH, and Q is N-Ph-R¹in Formula I and III, R¹ of Formula I and III is not selected from thegroup of halogen, alkyl, substituted alkyl, carboxylic acid, andcarboxylic esters.

Certain embodiments include a compound having the structure of Formula(Ia):

a tautomer thereof, or a pharmaceutically acceptable salt thereof,wherein:

R¹ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₃-C₆ cycloalkyl, an optionally substituted C₃-C₆cycloalkenyl, an optionally substituted C₂-C₆ heterocyclyl, anoptionally substituted aryl, and an optionally substituted heteroaryl;

R² and R³ are independently selected from hydrogen, an optionallysubstituted C₁-C₆ alkyl, an optionally substituted C₃-C₈ cycloalkyl, anoptionally substituted C₁-C₆ heterocycle, an optionally substitutedaryl, and an optionally substituted heteroaryl;

R⁴ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₃-C₈ cycloalkyl, an optionally substituted C₃-C₈cycloalkenyl, an optionally substituted C₁-C₆ heterocycle, an optionallysubstituted aryl, and an optionally substituted heteroaryl;

Q is selected from an optionally substituted C₁-C₆ alkyl, an optionallysubstituted C₃-C₈ cycloalkyl, and an optionally substituted non-aromaticheterocycle; and

Z is O (oxygen) or S (sulfur).

The compound of claim 2, wherein:

R¹ is selected from hydrogen, and an optionally substituted C₁-C₆ alkyl;

R² and R³ are independently selected from hydrogen, and an optionallysubstituted C₁-C₆ alkyl;

R⁴ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted aryl, and an optionally substituted heteroaryl;

Q is selected from an optionally substituted C₁-C₆ alkyl, and anoptionally substituted C₃-C₈ cycloalkyl; and

Z is O (oxygen).

The compound of claim 3, wherein:

R¹ is selected from hydrogen, and an optionally substituted C₁-C₃ alkyl;

R² and R³ are independently selected from hydrogen, and an optionallysubstituted C₁-C₃ alkyl;

R⁴ is selected from optionally substituted C₁-C₆ alkyl, and anoptionally substituted aryl; and

Q is selected from an optionally substituted C₁-C₃ alkyl, and anoptionally substituted C₃-C₃ cycloalkyl.

The compound of claim 4, wherein:

R¹ is hydrogen;

R² and R³ are independently an optionally substituted C₁-C₃ alkyl;

R⁴ is an optionally substituted phenyl; and

Q is an optionally substituted C₁-C₃ alkyl.

Certain embodiments include a compound having the structure of Formula(IIa) or (IIb):

a tautomer thereof, or a pharmaceutically acceptable salt thereof,wherein:

R³ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₃-C₈ cycloalkyl, an optionally substituted C₁-C₆heterocycle, an optionally substituted aryl, and an optionallysubstituted heteroaryl;

R⁴ is selected from an optionally substituted C₁-C₆ alkyl, an optionallysubstituted C₃-C₈ cycloalkyl, an optionally substituted C₁-C₆heterocyclyl, an optionally substituted aryl, an optionally substitutedheteroaryl, an optionally substituted arylalkyl, and an optionallysubstituted heteroarylalkyl; and

R⁹ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₃-C₈ cycloalkyl, an optionally substituted C₁-C₆heterocyclyl, an optionally substituted heteroaryl, an optionallysubstituted arylalkyl, and an optionally substituted heteroarylalkyl.

The compound of claim 6, wherein:

R³ is selected from hydrogen, and an optionally substituted C₁-C₃ alkyl;

R⁴ is selected from an optionally substituted C₁-C₆ alkyl, an optionallysubstituted C₃-C₈ cycloalkyl, and an optionally substituted aryl; and

R⁹ is selected from an optionally substituted C₁-C₆ alkyl, an optionallysubstituted arylalkyl, an optionally substituted arylalkenyl, anoptionally substituted arylalkynyl, and an optionally substitutedheteroarylalkyl.

In certain embodiments, R³ is an optionally substituted C₁-C₃ alkyl;

R⁴ is selected from an optionally substituted C₁-C₄ alkyl, and anoptionally substituted aryl; and

R⁹ is selected from an optionally substituted C₁-C₃ alkyl, an optionallysubstituted arylalkyl, an optionally substituted arylalkenyl, and anoptionally substituted arylalkynyl.

Certain embodiments include a compound having the structure of Formula(IIIa):

a tautomer thereof, or a pharmaceutically acceptable salt thereof,wherein:

R¹ is selected from hydrogen, OR⁶, NR⁶R⁷, CO₂R⁶, C(═O)NR⁶R⁷, anoptionally substituted C₂-C₆ heterocyclyl, an optionally substitutedarylalkyl, an optionally substituted aryl, and an optionally substitutedheteroaryl;

R² is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₃-C₈ cycloalkyl, an optionally substituted aryl,and an optionally substituted heteroaryl;

R⁴ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted arylalkyl, and an optionally substitutedheteroarylalkyl;

R⁵ is selected from hydrogen, halogen, CN, CF₃, OR⁶, an optionallysubstituted aryl, and an optionally substituted C₁-C₆ alkyl;

R⁶ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted aryl, and an optionally substituted heteroaryl;

R⁷ is selected from hydrogen, C(═O)R⁸, C(═O)NHR⁸, and an optionallysubstituted C₁-C₆ alkyl; or —NR⁶R⁷ is an optionally substitutednon-aromatic heterocycle linked through a ring nitrogen;

R⁸ is selected from hydrogen, and an optionally substituted C₁-C₆ alkyl;

Q is selected from NR⁶, an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₃-C₈ cycloalkyl, and an optionally substitutednon-aromatic heterocyclyl;

Z is O (oxygen) or S (sulfur); and

n is 1, or 2.

In certain embodiments, wherein R¹ is selected from hydrogen, OR⁶,NR⁶R⁷, CO₂R⁶, C(═O)NR⁶R⁷, an optionally substituted C₂-C₆ heterocyclyl,an optionally substituted arylalkyl, an optionally substituted aryl, andan optionally substituted heteroaryl;

R² is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₃-C₈ cycloalkyl, an optionally substituted aryl,and an optionally substituted heteroaryl;

R⁴ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted arylalkyl, and an optionally substitutedheteroarylalkyl;

R⁵ is selected from hydrogen, halogen, CN, CF₃, OR⁶, an optionallysubstituted aryl, and an optionally substituted C₁-C₆ alkyl;

R⁶ is selected from hydrogen, and an optionally substituted C₁-C₆ alkyl;

R⁷ is selected from hydrogen, C(═O)R⁸, C(═O)NHR⁸, and an optionallysubstituted C₁-C₆ alkyl;

R⁸ is selected from hydrogen, and an optionally substituted C₁-C₆ alkyl;

Q is selected from NR⁶, an optionally substituted C₁-C₄ alkyl, anoptionally substituted C₃-C₆ cycloalkyl, and an optionally substitutednon-aromatic heterocyclyl;

Z is O (oxygen); and

n is 1.

In certain embodiments, R¹ is selected from hydrogen, OR⁶, NR⁶R⁷,C(═O)NR⁶R⁷, an optionally substituted arylalkyl, and an optionallysubstituted heteroaryl;

R² is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted aryl, and an optionally substituted heteroaryl;

R⁴ is an optionally substituted aryl;

R⁵ is selected from hydrogen, chloro, CN, CF₃, OR⁶, an optionallysubstituted aryl, and an optionally substituted C₁-C₆ alkyl;

R⁶ is selected from hydrogen, and an optionally substituted C₁-C₃ alkyl;

R⁷ is selected from hydrogen, C(═O)R⁸, C(═O)NHR⁸, and an optionallysubstituted C₁-C₃ alkyl;

R⁸ is selected from hydrogen, and an optionally substituted C₁-C₃ alkyl;and

Q is selected from optionally substituted C₁-C₃ alkyl, an optionallysubstituted C₃-C₆ cycloalkyl, and an optionally substituted non-aromaticheterocyclyl.

Certain embodiments include a compound having the structure of Formula(IVa) or (IVb):

a tautomer thereof, or a pharmaceutically acceptable salt thereof,wherein:

R⁴ is selected from an optionally substituted C₁-C₆ alkyl, an optionallysubstituted C₃-C₈ cycloalkyl, an optionally substituted C₁-C₆heterocyclyl, an optionally substituted aryl, an optionally substitutedheteroaryl, an optionally substituted arylalkyl, and an optionallysubstituted heteroarylalkyl;

R⁵ is selected from halogen, CN, CF₃, OR⁶, an optionally substitutedaryl, and an optionally substituted C₁-C₆ alkyl;

R⁶ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted aryl, and an optionally substituted heteroaryl;and

R⁹ is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₃-C₈ cycloalkyl, an optionally substituted C₁-C₆heterocyclyl, an optionally substituted heteroaryl, an optionallysubstituted arylalkyl, and an optionally substituted heteroarylalkyl.

In certain embodiments, R⁴ is selected from an optionally substitutedC₁-C₆ alkyl, an optionally substituted C₃-C₈ cycloalkyl, and anoptionally substituted aryl;

R⁵ is selected from chloro, CN, CF₃, OR⁶, an optionally substitutedaryl, and an optionally substituted C₁-C₆ alkyl;

R⁶ is selected from hydrogen, and an optionally substituted C₁-C₃ alkyl;

R⁹ is selected from an optionally substituted C₁-C₆ alkyl, an optionallysubstituted arylalkyl, an optionally substituted arylalkenyl, anoptionally substituted arylalkynyl, and an optionally substitutedheteroarylalkyl, and

n is 1, or 2.

In certain embodiments, R⁴ is selected from an optionally substitutedC₁-C₄ alkyl, and an optionally substituted aryl;

R⁵ is selected from chloro, CN, CF₃, and an optionally substituted C₁-C₃alkyl;

R⁹ is selected from an optionally substituted C₁-C₃ alkyl, an optionallysubstituted arylalkyl, an optionally substituted arylalkenyl, and anoptionally substituted arylalkynyl; and

n is 1.

In certain embodiments, unless otherwise specified, groups indicated as“optionally substituted” are optionally substituted with one or moregroup(s) individually and independently selected from the groupconsisting of C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆heteroalkyl, C₁-C₆ haloalkyl, C₃-C₆ cycloalkyl, aryl, arylalkyl,alkenylO—, arylalkylO—, arylalkylNH—, alkenylO—, cycloalkylC(═O)—,arylC(═O)—, arylC(═O)NH—, arylNHC(═O)—, aryl(CH₂)₀₋₃O(CH₂)₀₋₃—,HO(CH₂)₁₋₃NH—, HO(CH₂)₁₋₃O—, HO(CH₂)₁₋₃—, HO(CH₂)₁₋₃O(CH₂)₁₋₃—,—C(═O)NHNH_(2,) heteroaryl, non-aromatic heterocycle, hydroxy, alkoxy,aryloxy, mercapto, alkylthio, arylthio, cyano, halo, oxo, O-carbamyl,N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido,S-sulfonamido, N-sulfonamido, C-carboxy, O-carboxy, nitro, and amino.

Certain embodiments include use of a compound for treating ahematopoietic or neurological disorder comprising administering aneffective amount of a compound of Formula (V) or (VI) to a subject inneed thereof, wherein Formula (V) or (VI):having the structure:

or pharmaceutically acceptable salt thereof, wherein:

R¹⁰ is selected from an optionally substituted C₁-C₁₂ alkyl, anoptionally substituted C₁-C₁₂ heteroalkyl, an optionally substitutedC₂-C₁₂ heteroalkenyl, an optionally substituted C₂-C₁₂ heteroalkynyl, anoptionally substituted C₁-C₁₂ cycloalkyl; an optionally substitutedC₁-C₆ heterocycloalkyl, an optionally substituted heteroarylalkyl, anoptionally substituted arylheteroalkyl, an optionally substitutedheteroarylheteroalkyl, OR¹⁴, SR¹⁴, and NR₈R¹⁴;

R¹¹ is selected from hydrogen, halogen, an optionally substituted C₁-C₆alkyl, an optionally substituted C₁-C₆ heteroalkyl, an optionallysubstituted C₃-C₈ cycloalkyl, and an optionally substituted C₁-C₆heterocycle;

R¹² is selected from an optionally substituted C₁-C₆ heteroalkyl, anoptionally substituted C₃-C₈ cycloalkyl, an optionally substituted C₁-C₆heterocycle, an optionally substituted aryl, and an optionallysubstituted heteroaryl;

R¹³ is selected from hydrogen, halogen, an optionally substituted C₁-C₈alkyl, an optionally substituted C₁-C₈ heteroalkyl, an optionallysubstituted C₂-C₈ heteroalkenyl, an optionally substituted C₃-C₈cycloalkyl, an optionally substituted C₁-C₆ heterocycloalkyl, OR¹⁴,SR¹⁴, and NR⁸R¹⁴;

R¹⁴ is selected from an optionally substituted aryl and an optionallysubstituted heteroaryl;

V is selected from CH═CH, N═CH, CH═N, NH, O (oxygen), and S (sulfur);and

n is 1, 2 or 3.

In certain embodiments, compounds as disclosed herein can exist intautomeric forms. All tautomeric forms are intended to be included inthe scope of the compounds as disclosed herein. Likewise, when compoundscontain a double bond, there exists the possibility of cis- andtrans-type isomeric forms of the compounds. Both cis- and trans-isomers,both in pure form as well as mixtures of cis- and trans-isomers, arecontemplated. Thus, reference herein to a compound includes all of theaforementioned isomeric forms unless the context clearly dictatesotherwise.

In certain embodiments, a compound of Formula (I) can reside in one ormore of the tautomeric forms. For example, the compound of Formula (I)can reside in the tautomeric forms shown below:

-   -   or their pharmaceutically acceptable salts.

In certain embodiments, a compound of Formula (II) can reside in one ormore of the tautomeric forms. For example, the compound of Formula (II)can reside in the tautomeric forms shown below:

-   -   or their pharmaceutically acceptable salts, wherein W can be NH;        and W¹ can be N (nitrogen).

In certain embodiments, a compound of Formula (III) can reside in one ormore of the tautomeric forms. For example, the compound of Formula (III)can reside in the tautomeric forms shown below:

-   -   or their pharmaceutically acceptable salts.

In certain embodiments, a compound of Formula (IV) can reside in one ormore of the tautomeric forms. For example, the compound of Formula (IV)can reside in the tautomeric forms shown below:

-   -   or their pharmaceutically acceptable salts, wherein W can be NH;        and W¹ can be N (nitrogen).

Certain Synthesis Methods

The process of Scheme I describes the general synthesis of compounds ofgeneral structure 3 described in Formula III, wherein R can be an alkylor heteroalkyl derivative. Treatment of the indolone derivatives ofgeneral structure 1 with a triethyl orthoester in acetic anhydrideaffords intermediates of general structure 2. The intermediates ofgeneral structure 2 are then condensed with a hydrazide to generate thecompounds of general structure 3.

The process of Scheme II describes general synthesis of the compounds ofFormula II. Compounds of structure 4 are diazotized under standardconditions and then are coupled with a ketoester of structure 5 toprovide hydrazone intermediates of structure 6. Condensation reaction ofthe intermediate ketoester of structure 6 and a hydrazine affords thefinal compounds of structure 7. Alternatively, compounds of structure 7can be obtained directly from coupling reaction of compounds ofstructure 8 with diazotized compounds of structure 4.

Scheme III describes general synthesis of compounds of Formula V and VI.The compounds of structure 11 can be prepared through a simple amideformation reaction between an acid derivative of structure 9 and a2-aminothiazole derivative of structure 10 with or without activation.

One of skill in the art will recognize that analogous synthesis schemesmay be used to synthesize compounds provided herein. One of skill willrecognize that compounds provided herein may be synthesized using othersynthesis schemes.

Certain Pharmaceutical Agents

In certain embodiments, a selective GCSFR modulator, or pharmaceuticallyacceptable salt, ester, amide, and/or prodrug thereof, either alone orcombined with one or more pharmaceutically acceptable carriers, forms apharmaceutical agent. Techniques for formulation and administration ofcompounds of the present embodiments may be found for example, in“Remington's Pharmaceutical Sciences,” Mack Publishing Co., Easton, Pa.,18th edition, 1990, which is incorporated herein by reference in itsentirety.

In certain embodiments, a pharmaceutical agent comprising one or morecompounds of the present embodiments is prepared using known techniques,including, but not limited to mixing, dissolving, granulating,dragee-making, levigating, emulsifying, encapsulating, entrapping ortabletting processes.

In certain embodiments, a pharmaceutical agent comprising one or morecompounds of the present embodiments is a liquid (e.g., a suspension,elixir and/or solution). In certain of such embodiments, a liquidpharmaceutical agent comprising one or more compounds of the presentembodiments is prepared using ingredients known in the art, including,but not limited to, water, glycols, oils, alcohols, flavoring agents,preservatives, and coloring agents.

In certain embodiments, a pharmaceutical agent comprising one or morecompounds of the present embodiments is a solid (e.g., a powder, tablet,and/or capsule). In certain of such embodiments, a solid pharmaceuticalagent comprising one or more compounds of the present embodiments isprepared using ingredients known in the art, including, but not limitedto, starches, sugars, diluents, granulating agents, lubricants, binders,and disintegrating agents.

In certain embodiments, a pharmaceutical agent comprising one or morecompounds of the present embodiments is formulated as a depotpreparation. Certain such depot preparations are typically longer actingthan non-depot preparations. In certain embodiments, such preparationsare administered by implantation (for example subcutaneously orintramuscularly) or by intramuscular injection. In certain embodiments,depot preparations are prepared using suitable polymeric or hydrophobicmaterials (for example an emulsion in an acceptable oil) or ion exchangeresins, or as sparingly soluble derivatives, for example, as a sparinglysoluble salt.

In certain embodiments, a pharmaceutical agent comprising one or morecompounds of the present embodiments comprises a delivery system.Examples of delivery systems include, but are not limited to, liposomesand emulsions. Certain delivery systems are useful for preparing certainpharmaceutical agents including those comprising hydrophobic compounds.In certain embodiments, certain organic solvents such asdimethylsulfoxide are used.

In certain embodiments, a pharmaceutical agent comprising one or morecompounds of the present embodiments comprises one or moretissue-specific delivery molecules designed to deliver thepharmaceutical agent to specific tissues or cell types. For example, incertain embodiments, pharmaceutical agents include liposomes coated witha tissue-specific antibody.

In certain embodiments, a pharmaceutical agent comprising one or morecompounds of the present embodiments comprises a co-solvent system.Certain of such co-solvent systems comprise, for example, benzylalcohol, a nonpolar surfactant, a water-miscible organic polymer, and anaqueous phase. In certain embodiments, such co-solvent systems are usedfor hydrophobic compounds. A non-limiting example of such a co-solventsystem is the VPD co-solvent system, which is a solution of absoluteethanol comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolarsurfactant Polysorbate 80™, and 65% w/v polyethylene glycol 300. Theproportions of such co-solvent systems may be varied considerablywithout significantly altering their solubility and toxicitycharacteristics. Furthermore, the identity of co-solvent components maybe varied: for example, other surfactants may be used instead ofPolysorbate 80™; the fraction size of polyethylene glycol may be varied;other biocompatible polymers may replace polyethylene glycol, e.g.,polyvinyl pyrrolidone; and other sugars or polysaccharides maysubstitute for dextrose.

In certain embodiments, a pharmaceutical agent comprising one or morecompounds of the present embodiments comprises a sustained-releasesystem. A non-limiting example of such a sustained-release system is asemi-permeable matrix of solid hydrophobic polymers. In certainembodiments, sustained-release systems may, depending on their chemicalnature, release compounds over a period of hours, days, weeks or months.

Certain compounds used in pharmaceutical agent of the presentembodiments may be provided as pharmaceutically acceptable salts withpharmaceutically compatible counterions. Pharmaceutically compatiblesalts may be formed with many acids, including but not limited tohydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic, etc.

In certain embodiments, a pharmaceutical agent comprising one or morecompounds of the present embodiments comprises an active ingredient in atherapeutically effective amount. In certain embodiments, thetherapeutically effective amount is sufficient to prevent, alleviate orameliorate symptoms of a disease or to prolong the survival of thesubject being treated. Determination of a therapeutically effectiveamount is well within the capability of those skilled in the art.

In certain embodiments, a pharmaceutical agent comprising one or morecompounds of the present embodiments is formulated as a prodrug. Incertain embodiments, prodrugs are useful because they are easier toadminister than the corresponding active form. For example, in certaininstances, a prodrug may be more bioavailable (e.g., through oraladministration) than is the corresponding active form. In certaininstances, a prodrug may have improved solubility compared to thecorresponding active form. In certain embodiments, a prodrug is anester. In certain embodiments, such prodrugs are less water soluble thanthe corresponding active form. In certain instances, such prodrugspossess superior transmittal across cell membranes, where watersolubility is detrimental to mobility. In certain embodiments, the esterin such prodrugs is metabolically hydrolyzed to carboxylic acid. Incertain instances the carboxylic acid containing compound is thecorresponding active form. In certain embodiments, a prodrug comprises ashort peptide (polyaminoacid) bound to an acid group. In certain of suchembodiments, the peptide is metabolized to form the corresponding activeform.

In certain embodiments, a pharmaceutical agent comprising one or morecompounds of the present embodiments is useful for treating a conditionsor disorder in a mammalian, and particularly in a human patient.Suitable administration routes include, but are not limited to, oral,rectal, transmucosal, intestinal, enteral, topical, suppository, throughinhalation, intrathecal, intraventricular, intraperitoneal, intranasal,intraocular and parenteral (e.g., intravenous, intramuscular,intramedullary, and subcutaneous). In certain embodiments,pharmaceutical intrathecals are administered to achieve local ratherthan systemic exposures. For example, pharmaceutical agents may beinjected directly in the area of desired effect (e.g., in the renal orcardiac area).

In certain embodiments, a pharmaceutical agent comprising one or morecompounds of the present embodiments is administered in the form of adosage unit (e.g., tablet, capsule, bolus, etc.). In certainembodiments, such dosage units comprise a selective HGF modulator in adose from about 1 μg/kg of body weight to about 50 mg/kg of body weight.In certain embodiments, such dosage units comprise a selective HGFmodulator in a dose from about 2 μg/kg of body weight to about 25 mg/kgof body weight. In certain embodiments, such dosage units comprise aselective HGF modulator in a dose from about 10 μg/kg of body weight toabout 5 mg/kg of body weight. In certain embodiments, pharmaceuticalagents are administered as needed, once per day, twice per day, threetimes per day, or four or more times per day. It is recognized by thoseskilled in the art that the particular dose, frequency, and duration ofadministration depends on a number of factors, including, withoutlimitation, the biological activity desired, the condition of thepatient, and tolerance for the pharmaceutical agent.

In certain embodiments, a pharmaceutical agent comprising a compound ofthe present embodiments is prepared for oral administration. In certainof such embodiments, a pharmaceutical agent is formulated by combiningone or more compounds of the present embodiments with one or morepharmaceutically acceptable carriers. Certain of such carriers enablecompounds of the present embodiments to be formulated as tablets, pills,dragees, capsules, liquids, gels, syrups, slurries, suspensions and thelike, for oral ingestion by a patient. In certain embodiments,pharmaceutical agents for oral use are obtained by mixing one or morecompounds of the present embodiments and one or more solid excipient.Suitable excipients include, but are not limited to, fillers, such assugars, including lactose, sucrose, mannitol, or sorbitol; cellulosepreparations such as, for example, maize starch, wheat starch, ricestarch, potato starch, gelatin, gum tragacanth, methyl cellulose,hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/orpolyvinylpyrrolidone (PVP). In certain embodiments, such a mixture isoptionally ground and auxiliaries are optionally added. In certainembodiments, pharmaceutical agents are formed to obtain tablets ordragee cores. In certain embodiments, disintegrating agents (e.g.,cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a saltthereof, such as sodium alginate) are added.

In certain embodiments, dragee cores are provided with coatings. Incertain of such embodiments, concentrated sugar solutions may be used,which may optionally contain gum arabic, talc, polyvinyl pyrrolidone,carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquersolutions, and suitable organic solvents or solvent mixtures. Dyestuffsor pigments may be added to tablets or dragee coatings.

In certain embodiments, pharmaceutical agents for oral administrationare push-fit capsules made of gelatin. Certain of such push-fit capsulescomprise one or more compounds of the present embodiments in admixturewith one or more filler such as lactose, binders such as starches,and/or lubricants such as talc or magnesium stearate and, optionally,stabilizers. In certain embodiments, pharmaceutical agents for oraladministration are soft, sealed capsules made of gelatin and aplasticizer, such as glycerol or sorbitol. In certain soft capsules, oneor more compounds of the present embodiments are be dissolved orsuspended in suitable liquids, such as fatty oils, liquid paraffin, orliquid polyethylene glycols. In addition, stabilizers may be added.

In certain embodiments, pharmaceutical agents are prepared for buccaladministration. Certain of such pharmaceutical agents are tablets orlozenges formulated in conventional manner.

In certain embodiments, a pharmaceutical agent is prepared foradministration by injection (e.g., intravenous, subcutaneous,intramuscular, etc.). In certain of such embodiments, a pharmaceuticalagent comprises a carrier and is formulated in aqueous solution, such aswater or physiologically compatible buffers such as Hanks's solution,Ringer's solution, or physiological saline buffer. In certainembodiments, other ingredients are included (e.g., ingredients that aidin solubility or serve as preservatives). In certain embodiments,injectable suspensions are prepared using appropriate liquid carriers,suspending agents and the like. Certain pharmaceutical agents forinjection are presented in unit dosage form, e.g., in ampoules or inmulti-dose containers. Certain pharmaceutical agents for injection aresuspensions, solutions or emulsions in oily or aqueous vehicles, and maycontain formulatory agents such as suspending, stabilizing and/ordispersing agents. Certain solvents suitable for use in pharmaceuticalagents for injection include, but are not limited to, lipophilicsolvents and fatty oils, such as sesame oil, synthetic fatty acidesters, such as ethyl oleate or triglycerides, and liposomes. Aqueousinjection suspensions may contain substances that increase the viscosityof the suspension, such as sodium carboxymethyl cellulose, sorbitol, ordextran. Optionally, such suspensions may also contain suitablestabilizers or agents that increase the solubility of the compounds toallow for the preparation of highly concentrated solutions.

In certain embodiments, a pharmaceutical agent is prepared fortransmucosal administration. In certain of such embodiments penetrantsappropriate to the barrier to be permeated are used in the formulation.Such penetrants are generally known in the art.

In certain embodiments, a pharmaceutical agent is prepared foradministration by inhalation. Certain of such pharmaceutical agents forinhalation are prepared in the form of an aerosol spray in a pressurizedpack or a nebulizer. Certain of such pharmaceutical agents comprise apropellant, e.g., dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas. Incertain embodiments using a pressurized aerosol, the dosage unit may bedetermined with a valve that delivers a metered amount. In certainembodiments, capsules and cartridges for use in an inhaler orinsufflator may be formulated. Certain of such formulations comprise apowder mixture of a compound of the present embodiments and a suitablepowder base such as lactose or starch.

In certain embodiments, a pharmaceutical agent is prepared for rectaladministration, such as a suppositories or retention enema. Certain ofsuch pharmaceutical agents comprise known ingredients, such as cocoabutter and/or other glycerides.

In certain embodiments, a pharmaceutical agent is prepared for topicaladministration. Certain of such pharmaceutical agents comprise blandmoisturizing bases, such as ointments or creams. Exemplary suitableointment bases include, but are not limited to, petrolatum, petrolatumplus volatile silicones, lanolin and water in oil emulsions such asEucerin™, available from Beiersdorf (Cincinnati, Ohio). Exemplarysuitable cream bases include, but are not limited to, Nivea™ Cream,available from Beiersdorf (Cincinnati, Ohio), cold cream (USP), PurposeCream™, available from Johnson & Johnson (New Brunswick, N.J.),hydrophilic ointment (USP) and Lubriderm™, available from Pfizer (MorrisPlains, N.J.).

In certain embodiments, the formulation, route of administration anddosage for a pharmaceutical agent of the present embodiments can bechosen in view of a particular patient's condition. (See e.g., Fingl etal. 1975, in “The Pharmacological Basis of Therapeutics”, Ch. 1 p. 1,which is incorporated herein by reference in its entirety). In certainembodiments, a pharmaceutical agent is administered as a single dose. Incertain embodiments, a pharmaceutical agent is administered as a seriesof two or more doses administered over one or more days.

In certain embodiments, a pharmaceutical agent of the presentembodiments is administered to a patient between about 0.1% and 500%, 5%and 200%, 10% and 100%, 15% and 85%, 25% and 75%, or 40% and 60% of anestablished human dosage. Where no human dosage is established, asuitable human dosage may be inferred from ED₅₀ or ID₅₀ values, or otherappropriate values derived from in vitro or in vivo studies.

In certain embodiments, a daily dosage regimen for a patient comprisesan oral dose of between 0.1 mg and 2000 mg, 5 mg and 1500 mg, 10 mg and1000 mg, 20 mg and 500 mg, 30 mg and 200 mg, or 40 mg and 100 mg of acompound of the present embodiments. In certain embodiments, a dailydosage regimen is administered as a single daily dose. In certainembodiments, a daily dosage regimen is administered as two, three, four,or more than four doses.

In certain embodiments, a pharmaceutical agent of the presentembodiments is administered by continuous intravenous infusion. Incertain of such embodiments, from 0.1 mg to 500 mg of a composition ofthe present embodiments is administered per day.

In certain embodiments, a pharmaceutical agent of the presentembodiments is administered for a period of continuous therapy. Forexample, a pharmaceutical agent of the present embodiments may beadministered over a period of days, weeks, months, or years.

Dosage amount, interval between doses, and duration of treatment may beadjusted to achieve a desired effect. In certain embodiments, dosageamount and interval between doses are adjusted to maintain a desiredconcentration on compound in a patient. For example, in certainembodiments, dosage amount and interval between doses are adjusted toprovide plasma concentration of a compound of the present embodiments atan amount sufficient to achieve a desired effect. In certain of suchembodiments the plasma concentration is maintained above the minimaleffective concentration (MEC). In certain embodiments, pharmaceuticalagents of the present embodiments are administered with a dosage regimendesigned to maintain a concentration above the MEC for 10-90% of thetime, between 30-90% of the time, or between 50-90% of the time.

In certain embodiments in which a pharmaceutical agent is administeredlocally, the dosage regimen is adjusted to achieve a desired localconcentration of a compound of the present embodiments.

In certain embodiments, a pharmaceutical agent may be presented in apack or dispenser device which may contain one or more unit dosage formscontaining the active ingredient. The pack may for example comprisemetal or plastic foil, such as a blister pack. The pack or dispenserdevice may be accompanied by instructions for administration. The packor dispenser may also be accompanied with a notice associated with thecontainer in form prescribed by a governmental agency regulating themanufacture, use, or sale of pharmaceuticals, which notice is reflectiveof approval by the agency of the form of the drug for human orveterinary administration. Such notice, for example, may be the labelingapproved by the U.S. Food and Drug Administration for prescriptiondrugs, or the approved product insert. Compositions comprising acompound of the present embodiments formulated in a compatiblepharmaceutical carrier may also be prepared, placed in an appropriatecontainer, and labeled for treatment of an indicated condition.

In certain embodiments, a pharmaceutical agent is in powder form forconstitution with a suitable vehicle, e.g., sterile pyrogen-free water,before use.

Certain Combination Therapies

In certain embodiments, one or more pharmaceutical agents of the presentembodiments are co-administered with one or more other pharmaceuticalagents. In certain embodiments, such one or more other pharmaceuticalagents are designed to treat the same disease or condition as the one ormore pharmaceutical agents of the present embodiments. In certainembodiments, such one or more other pharmaceutical agents are designedto treat a different disease or condition as the one or morepharmaceutical agents of the present embodiments. In certainembodiments, such one or more other pharmaceutical agents are designedto treat an undesired effect of one or more pharmaceutical agents of thepresent embodiments. In certain embodiments, one or more pharmaceuticalagents of the present embodiments are co-administered with anotherpharmaceutical agent to treat an undesired effect of that otherpharmaceutical agent. In certain embodiments, one or more pharmaceuticalagents of the present embodiments and one or more other pharmaceuticalagents are administered at the same time. In certain embodiments, one ormore pharmaceutical agents of the present embodiments and one or moreother pharmaceutical agents are administered at the different times. Incertain embodiments, one or more pharmaceutical agents of the presentembodiments and one or more other pharmaceutical agents are preparedtogether in a single formulation. In certain embodiments, one or morepharmaceutical agents of the present embodiments and one or more otherpharmaceutical agents are prepared separately.

Examples of pharmaceutical agents that may be co-administered with apharmaceutical agent of the present embodiments include, but are notlimited to, anti-cancer treatments, including, but not limited to,chemotherapy and radiation treatment; corticosteroids, including but notlimited to prednisone; immunoglobulins, including, but not limited tointravenous immunoglobulin (IVIg); analgesics (e.g., acetaminophen);anti-inflammatory agents, including, but not limited to non-steroidalanti-inflammatory drugs (e.g., ibuprofen, COX-1 inhibitors, and COX-2,inhibitors); salicylates; antibiotics; antivirals; antifungal agents;antidiabetic agents (e.g., biguanides, glucosidase inhibitors, insulins,sulfonylureas, and thiazolidenediones); adrenergic modifiers; diuretics;hormones (e.g., anabolic steroids, androgen, estrogen, calcitonin,progestin, somatostatin, and thyroid hormones); immunomodulators; musclerelaxants; antihistamines; osteoporosis agents (e.g., biphosphonates,calcitonin, and estrogens); prostaglandins, antineoplastic agents;psychotherapeutic agents; sedatives; poison oak or poison sumacproducts; antibodies; and vaccines.

In some embodiments, the compounds provided herein can be administeredin combination with an additional therapeutic regimen. In some suchembodiments, the additional therapeutic regimen can includechemotherapy, bone marrow transplantation, and radiation therapy. Incertain embodiments, a compound provided herein can be administered to asubject in combination with harvesting peripheral blood progenitor cellsand/or in conjunction with hematopoietic stem cell transplantation. Suchadministration may be done before, during, and/or after such harvesting.

Certain Indications

Certain embodiments include methods of treating or ameliorating adisorder such as a hematopoietic or neurological disorder. Some suchmethods can include administering an effective amount of a compoundprovided herein to a subject in need thereof. Examples of disorders thatmay be treated with the compounds provided herein includegranulocytopenia, neutropenia, amyotrophic lateral sclerosis, multiplesclerosis, multiple dystrophy, injury to the nervous system, such asspinal cord injury and injury resulting from trauma or from stroke. Incertain embodiments, a disorder can include granulocytopenia.Granulocytopenia can result from chemotherapy, radiation treatment,failure of a bone marrow transplantation, and/or aplastic anemia.

In certain embodiments, the compounds provided herein can be useful topromote growth and/or development of glial cells. Such glial cells mayrepair nerve cells. In certain embodiments, compounds provided hereinare useful to treat psychological disorders, including, but not limitedto, cognitive disorders.

In certain embodiments, the compounds provided herein can be useful totreat disorders associated with abnormal function of hematopoiesis,erythropoiesis, granulopoiesis, thrombopoiesis, and myelopoiesis.Examples of such disorders include anemia, neutropenia,thrombocytopenia, cardiovascular disorders, immune/autoimmune disorders,cancers, infectious disorders or diseases, and neurologic disorders. Insome embodiments, the compounds provided herein can be administered withadditional agents useful to modulate hematopoiesis, erythropoiesis,granulopoiesis, thrombopoiesis, and myelopoiesis.

Certain Methods of Identifying Compounds

Some embodiments of the methods provided herein include identifyingtherapeutic compounds. Some such therapeutic compounds include compoundsthat stimulate granulopoiesis, such as GCSFR agonists. Some embodimentsinclude identifying compounds that selectively modulate the activity ofa GCSFR. In some embodiments, therapeutic compounds include compoundswith reduced activity in the presence of a mutant GCSFR protein comparedto a wild type GCSFR protein. In some such embodiments, the mutant GCSFRcomprises a mutation in the transmembrane domain of the GCSFR protein,or a mutation proximal to the transmembrane domain of the GCSFR protein.

Some methods for identifying a therapeutic compound include contacting atarget cell with a test compound comprising a GCSF receptor agonist,wherein the target cell comprises a mutant GCSF receptor protein; anddetermining whether the test compound significantly changes the level ofactivity of the mutant GCSF receptor in the target cell. Someembodiments also include comparing the level of activity of the mutantGCSF receptor in the target cell to the level of activity of a wild typeGCSF receptor protein in a cell contacted with the test compound. Someembodiments also include determining whether the level of activity ofthe mutant GCSF receptor in the target cell contacted with the testcompound is less than the level of activity of the wild type GCSFreceptor protein in a cell contacted with the test compound. In someembodiments, the cell comprises a mammalian cell. In some embodiments,the cell comprises a human cell. In some embodiments, the cell is not ablood cell.

In some embodiments, the mutant GCSF receptor protein comprises amutation in the transmembrane domain of the protein or a mutationproximal to the transmembrane domain of the protein. The mutation caninclude a substitution, deletion or insertion. In some such embodiments,the mutation can include a substitution at the residue corresponding tohis-627 of the human GCSF receptor protein, and a substitution at theresidue corresponding to Asp-602 of the mouse GCSF receptor protein. Insome embodiments the mutant GCSFR includes a substitution at a residuecorresponding to a residue proximal to the TM domain of the polypeptide,for example, the substituted residue can be no more than 1, 2, 3, 4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24,25, 26, 27, 28, 29, and 30 consecutive residues from the TM domain ofthe GCSFR polypeptide. In some embodiments, the mutant GCSF receptorcomprises a mutant human GCSF receptor. An example polypeptide sequence(SEQ ID No:01) of a human GCSF receptor protein is shown in Table 1.

TABLE 1Granulocyte colony-stimulating factor isoform d precursor [Homo sapiens].ACCESSION NP_001171618   1magpatqspm klmalqlllw hsalwtvqea tplgpasslp qsfllkcleq vrkiqgdgaa  61lqeklagcls qlhsglflyq gllqalegis pelgptldtl qldvadfatt iwqqmeelgm 121apalqptqga mpafasafqr raggvlvash lqsflevsyr vlrhlaqp

In more methods of identifying a therapeutic compound, a cell iscontacted with a test compound and the level of activity of the GCSFR isdetermined. In some embodiments, the level of activity of the GCSFR in acell contacted with the test compound is compared to the level ofactivity of the GCSFR in a cell not contacted with the test compound. Insome embodiments, an increase in the activity of the GCSFR in a cellcontacted with a test compound compared to the activity in the GCSFR ina cell not contacted with the test compound is indicative that the testcompound activates GCSFR. The level of activity of GCSFR can be measuredby a variety of methods well known in the art.

Some embodiments for identifying compounds that selectively modulate theactivity of a GCSFR can include identifying compounds that increase theactivity of a wild type TPO receptor (TPOR) compared to a mutant TPOR.In some embodiments, the mutant TPOR includes a substitution at theresidue corresponding to his-499 of the human TPOR polypeptide. In someembodiments, a compound increases the activity of a human TPOR but not amouse TPOR or monkey TPOR.

In certain embodiments, methods of screening small molecule compoundlibrary using the luciferase reporter assays are employed to identifythe binding-site selective hGCSFR activator compounds. In certainembodiments, the compound library includes small molecule compounds thatactivate TPOR by requiring histidine-499 of human TPOR (hTPOR) TM domainor that selectively activate hTPOR but not mouse and monkey TPOreceptors. In certain embodiments, the [¹²⁵I]-rhGCSF competitivebinding, cell proliferation, and bone marrow cell differential assaysare used to characterize the small molecule binding-site selectivehGCSFR activator compounds.

EXAMPLES Example 1

4-Amino-4-oxobutyric acidN′-(phenyl-1-(1-(3,5-dimethoxyphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)methyl)hydrazide(Compound 101)

Compound 101 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethoxyphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthobenzoate, and 4-amino-4-oxobutyric acid hydrazide. ¹H NMR(400 MHz, MeOD) 7.62 (m, 3H), 7.44 (m, 2H), 6.92 (m, 2H), 6.62 (m, 3H),5.98 (m, 1H), 3.82 (m, 6H), 2.28 (m, 2H), 2.20 (m, 2H). MW=554, LC-MS(M+1)=554.98.

Example 2

4-Amino-4-oxobutyric acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)pentyl)hydrazide(Compound 102)

Compound 102 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthovalerate, and 4-amino-4-oxobutyric acid hydrazide. ¹H NMR(400 MHz, MeOD) 7.53 (d, J=7.5, 1H), 7.38 (d, J=7.5, 1H), 7.18 (s, 1H),7.05 (s, 1H), 6.94 (s, 1H), 2.95 (m, 2H), 2.62 (s, 4H), 2.42 (m, 6H),1.73 (m, 2H), 1.65 (m, 2H), 1.05 (t, J=8.0, 3H). MW=502, LC-MS(M+1)=502.99.

Example 3

4-Amino-4-oxobutyric acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)butyl)hydrazide(Compound 103)

Compound 103 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthobutyrate, and 4-amino-4-oxobutyric acid hydrazide. ¹H NMR(400 MHz, MeOD) 7.52 (d, J=7.5, 1H), 7.36 (d, J=7.5, 1H), 7.16 (s, 1H),7.02 (s, 2H), 6.91 (s, 1H), 2.89 (m, 2H), 2.62 (s, 4H), 2.39 (s, 6H),1.78 (m, 2H), 1.19 (t, J=7.0, 3H). MW=488, LC-MS (M+1)=488.95.

Example 4

Cyclopentanecarboxylic acidN′-(1-phenyl-1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)methyl)hydrazide(Compound 104)

Compound 104 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthobenzoate, and cyclopentylcarboxylic acid hydrazide.MW=519, LC-MS (M+1)=519.98.

Example 5

3-Acetylaminopropionic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 105)

Compound 105 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 3-acetylaminopropionic acid hydrazide. ¹H NMR(400 MHz, MeOD) 7.66 (d, J=7.5, 1H), 7.35 (d, J=7.5, 1H), 7.16 (s, 1H),7.02 (s, 2H), 6.92 (s, 1H), 3.50 (m, 2H), 2.55 (s, 5H), 2.40 (s, 6H),1.95 (s, 3H). MW=474, LC-MS (M+1)=475.01.

Example 6

4-Aminobutyric acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 106)

Compound 106 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 4-aminobutyric acid hydrazide.

Example 7

(5-Amino-1-tetrazole)acetic acidN′-(1-phenyl-1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)methyl)hydrazide(Compound 107)

Compound 107 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthobenzoate, and 2-(5-amino-1-tetrazole)acetic acidhydrazide. MW=548, LC-MS (M+1)=549.01.

Example 8

(1-Tetrazole)acetic acidN′-(1-phenyl-1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)methyl)hydrazide(Compound 108)

Compound 108 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthobenzoate, and 1-tetrazoleacetic acid hydrazide. MW=533,LC-MS (M+1)=533.96.

Example 9

(±)-(1-Benzyl-2-oxo-5-pyrrolidine)carboxylic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 109)

Compound 109 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and (1-benzyl-2-oxo-4-pyrrolidine)carboxylic acidhydrazide. MW=562, LC-MS (M+1)=563.05.

Example 10

4-Amino-4-oxobutyric acidN′-(1-(1-(3-methoxyphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 110)

Compound 110 was prepared according to the procedure described in SchemeI from 1-(3-methoxyphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 4-amino-4-oxobutyric acid hydrazide. ¹H NMR(400 MHz, MeOD) 7.67 (m, 1H), 7.50 (m, 1H), 7.35 (m, 1H), 7.06 (m, 1H),6.98 (m, 3H), 3.85 (m, 3H), 2.60 (m, 7H). MW=462, LC-MS (M+1)=463.02.

Example 11

4-Amino-4-oxobutyric acidN′-(1-(1-(3,5-dimethoxyphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 111)

Compound 111 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethoxyphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 4-amino-4-oxobutyric acid hydrazide. ¹H NMR(400 MHz, MeOD) 7.59 (m, 1H), 7.35 (m, 1H), 7.03 (m, 1H), 6.60 (m, 3H),3.83 (m, 3H), 2.62 (m, 7H). MW=492, LC-MS (M+1)=492.99.

Example 12

4-Dimethylamino-4-oxobutyric acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 112)

Compound 112 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 4-dimethylamino-4-oxobutyric acid hydrazide.¹H NMR (400 MHz, MeOD) 7.65 (d, J=7.5, 1H), 7.34 (d, J=7.5, 1H), 7.14(s, 1H), 7.01 (s, 2H), 6.92 (s, 1H), 3.08 (s, 3H), 2.95 (s, 3H), 2.77(m, 2H), 2.63 (m, 2H), 2.57 (s, 3H), 2.39 (s, 6H). MW=488, LC-MS(M+1)=489.07.

Example 13

4-Hydroxycyclohexyl-1-carboxylic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 113)

Compound 113 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 4-hydroxycyclohexylcarboxylic acid hydrazide.MW=487, LC-MS (M+1)=487.99.

Example 14

5-Hydroxy-5-oxopentanoic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 114)

Compound 114 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 5-hydroxy-5-oxovaleric acid hydrazide.MW=475, LC-MS (M+1)=475.98.

Example 15

3-Amino-3-oxopropionic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 115)

Compound 115 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 3-amino-3-oxopropionic acid hydrazide.MW=446, LC-MS (M+1)=446.89.

Example 16

(±)-(2-Oxo-3-piperidine)carboxylic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 116)

Compound 116 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 2-oxopiperidinecarboxylic acid hydrazide.MW=486, LC-MS (M+1)=486.98.

Example 17

4-Amino-4-oxobutyric acidN′-(1-phenyl-1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)methyl)hydrazide(Compound 117)

Compound 117 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthobenzoate, and 4-amino-4-oxobutyric acid hydrazide. 1H NMR(400 MHz, MeOD) 7.63 (m, 3H), 7.45 (d, J=7.5), 7.17 (s, 1H), 7.05 (s,1H), 6.90 (d, J=7.5, 1H), 6.84 (s, 1H), 5.98 (s, 1H), 2.41 (s, 6H), 2.27(m, 2H), 2.21 (m, 2H). MW=522, LC-MS (M+1)=523.02.

Example 18

3-(1-Pyrazole)propionic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 118)

Compound 118 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 3-(1-pyrazole)propionic acid hydrazide.MW=483, LC-MS (M+1)=484.01.

Example 19

4-Hydroxy-4-oxobutyric acidN′-(1-(1-(4-methylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 119)

Compound 119 was prepared according to the procedure described in SchemeI from 1-(4-methylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and succinic acid monohydrazide. 1H NMR (400 MHz,MeOD) 7.66 (m, 1H), 7.42 (m, 2H), 7.35 (m, 1H), 7.30 (m, 2H), 6.92 (m,1H), 2.67 (m, 2H), 2.61 (m, 2H), 2.58 (m, 3H), 2.46 (m, 3H). MW=447,LC-MS (M+1)=447.97.

Example 20

4-Amino-4-oxobutyric acidN′-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)methylhydrazide(Compound 120)

Compound 120 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthocarboxylate, and 4-amino-4-oxobutyric acid hydrazide. ¹HNMR (400 MHz, MeOD) 7.58 (m, 1H), 7.31 (m. 1H), 7.15 (s, 1H), 7.04 (s,2H), 6.90 (s, 1H), 2.61 (m, 4H), 2.41 (s, 6H). MW=446, LC-MS(M+1)=446.96.

Example 21

4-Amino-4-oxobutyric acidN′-(1-(1-(4-methylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 121)

Compound 121 was prepared according to the procedure described in SchemeI from 1-(4-methylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 4-amino-4-oxobutyric acid hydrazide. ¹H NMR(400 MHz, MeOD) 7.66 (d, J=7.5, 1H), 7.42 (d, J=7.5, 2H), 7.35 (d,J=7.5, 1H), 7.28 (d, J=7.5, 2H), 6.93 (s, 1H), 2.62 (m, 4H), 2.57 (s,3H), 2.46 (s, 3H). MW=446, LC-MS (M+1)=446.96.

Example 22

Cyclopropylcarboxylic acidN′-(1-(1-(3-methylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 122)

Compound 122 was prepared according to the procedure described in SchemeI from 1-(3-methylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and cyclopropylcarboxylic acid hydrazide. ¹H NMR(400 MHz, MeOD) 7.66 (m, 1H), 7.46 (m, 1H), 7.34 (m, 2H), 7.22 (m, 2H),6.92 (s, 1H), 2.55 (s, 3H), 2.43 (s, 3H), 1.71 (m, 1H), 0.95 (m, 4H).MW=415, LC-MS (M+1)=415.99.

Example 23

4-Hydroxybutyric acidN′-(1-phenyl-1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)methyl)hydrazide(Compound 123)

Compound 123 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthobenzoate, and 4-hydroxybutyric acid hydrazide. MW=509,LC-MS (M+1)=509.99.

Example 24

Cyclopropylcarboxylic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)-1-(2-furyl)methyl)hydrazide(Compound 124)

Compound 124 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl ortho-(2-furano)carboxylate, and cyclopropylcarboxylic acidhydrazide.

Example 25

Cyclopropylcarboxylic acidN′-(1-(1-(4-methylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 125)

Compound 125 was prepared according to the procedure described in SchemeI from 1-(4-methylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and cyclopropylcarboxylic acid hydrazide.1H NMR(400 MHz, MeOD) 7.65 (d, J=7.5, 1H), 7.42 (d, J=7.5, 2H), 7.35 (d,J=7.5, 1H), 7.28 (d, J=7.5, 2H), 6.92 (s, 1H), 2.55 (s, 3H), 2.45 (s,3H), 1.71 (m, 1H), 0.95 (m, 4H). MW=415, LC-MS (M+1)=415.99.

Example 26

4-Hydroxybutyric acidN′-(1-(1-(3-trifluoromethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 126)

Compound 126 was prepared according to the procedure described in SchemeI from1-(3-trifluoromethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 4-hydroxybutyric acid hydrazide. MW=487,LC-MS (M+1)=487.99.

Example 27

Butyric acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 127)

Compound 127 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and butyric acid hydrazide. MW=431, LC-MS(M+1)=431.98.

Example 28

4-Amino-4-oxobutyric acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 128)

Compound 128 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 4-amino-4-oxobutyric acid hydrazide. MW=460,LC-MS (M+1)=461.00.

Example 29

4-Phenylamino-4-oxobutyric acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 129)

Compound 129 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 4-phenylamino-4-oxobutyric acid hydrazide.MW=536, LC-MS (M+1)=537.06.

Example 30

4-Hydroxybutyric acidN′-(1-(1-(4-methylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 130)

Compound 130 was prepared according to the procedure described in SchemeI from 1-(4-methylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 4-hydroxybutyric acid hydrazide. MW=433,LC-MS (M+1)=433.94.

Example 31

4-Hydroxybutyric acidN′-(1-(1-(3-methylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 131)

Compound 131 was prepared according to the procedure described in SchemeI from 1-(3-methylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 4-hydroxybutyric acid hydrazide. MW=433,LC-MS (M+1)=433.94.

Example 32

Cyclopropylcarboxylic acidN′-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)methylhydrazide(Compound 132)

Compound 132 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthocarboxylate, and cyclopropylcarboxylic acid hydrazide.

Example 33

3-Methoxypropionic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 133)

Compound 133 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 3-methoxypropionic acid hydrazide. MW=447,LC-MS (M+1)=447.98.

Example 34

3-Indoleacetic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 134)

Compound 134 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 3-indoleacetic acid hydrazide. MW=518, LC-MS(M+1)=519.04.

Example 35

4-Hydroxybutyric acidN′-(1-(1-phenyl-2-oxo-6-methoxy-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 135)

Compound 135 was prepared according to the procedure described in SchemeI from 1-phenyl-6-methoxy-1,2-dihydroindol-2-one, triethyl orthoacetate,and 4-hydroxybutyric acid hydrazide. MW=381, LC-MS (M+1)=382.00.

Example 36

4-Hydroxybutyric acidN′-(1-(1-phenyl-2-oxo-6-methyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 136)

Compound 136 was prepared according to the procedure described in SchemeI from 1-phenyl-6-methyl-1,2-dihydroindol-2-one, triethyl orthoacetate,and 4-hydroxybutyric acid hydrazide. MW=365, LC-MS (M+1)=366.01.

Example 37

4-Hydroxyvaleric acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 137)

Compound 137 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 4-hydroxyvaleric acid hydrazide. MW=461,LC-MS (M+1)=461.94.

Example 38

3-Hydroxypropionic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 138)

Compound 138 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 3-hydroxypropionic acid hydrazide. MW=433,LC-MS (M+1)=433.94.

Example 39

4-Hydroxybutyric acidN′-(1-(1-phenyl-2-oxo-6-cyano-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 139)

Compound 139 was prepared according to the procedure described in SchemeI from 1-phenyl-6-trifluoromethyl-1,2-dihydroindol-2-one, triethylorthoacetate, and 4-hydroxybutyric acid hydrazide. MW=376, LC-MS(M+1)=377.01.

Example 40

4-Hydroxybutyric acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)propyl)hydrazide(Compound 140)

Compound 140 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthopropionate, and 4-hydroxybutyric acid hydrazide. MW=461,LC-MS (M+1)=461.94.

Example 41

Aminocarboxylic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)propyl)hydrazide(Compound 141)

Compound 141 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthopropionate, and 1-aminocarboxylic acid hydrazide. MW=418,LC-MS (M+1)=418.96.

Example 42

4-Hydroxybutyric acidN′-(1-(1-phenyl-2-oxo-6-chloro-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 142)

Compound 142 was prepared according to the procedure described in SchemeI from 1-phenyl-6-chloro-1,2-dihydroindol-2-one, triethyl orthoacetate,and 4-hydroxybutyric acid hydrazide.

Example 43

Aminocarboxylic acidN′-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)methylhydrazide(Compound 143)

Compound 143 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthocarboxylate, and 1-aminocarboxylic acid hydrazide. MW=390,LC-MS (M+1)=390.97.

Example 44

4-Hydroxybutyric acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 144)

Compound 144 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 4-hydroxybutyric acid hydrazide. MW=447,LC-MS (M+1)=447.91.

Example 45

3-Dimethylaminopropionic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 145)

Compound 145 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 3-dimethylaminopropionic acid hydrazide.MW=460, LC-MS (M+1)=461.00.

Example 46

2-Cyanoacetic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 146)

Compound 146 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and cyanoacetic acid hydrazide. MW=428, LC-MS(M+1)=428.95.

Example 47

(±)-3-Hydroxybutyric acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 147)

Compound 147 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 3-hydroxybutyric acid hydrazide. 1H NMR (400MHz, DMSO-d₆) 11.55 (s, 1H), 10.56 (s, 1H), 7.70 (d, J=8.0, 1H), 7.38(d, J=8.0, 1H), 7.12 (s, 1H), 7.06 (s, 1H), 6.89 (s, 1H), 4.88 (d,J=8.0, 1H), 4.39 (t, J=5.0 and 8.0, 1H), 4.05 (m, 2H), 2.58 (s, 3H),2.49 (s, 6H), 2.32 (d, J=8.0, 3H).

Example 48

Cyclopropylcarboxylic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 148)

Compound 148 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and cyclopropylcarboxylic acid hydrazide. MW=429,LC-MS (M+1)=429.89.

Example 49

Aminothiocarboxylic acidN′-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)methylhydrazide(Compound 149)

Compound 149 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthocarboxylate, and 1-aminothiocarboxylic acid hydrazide.MW=420, LC-MS (M+1)=420.92.

Example 50

Acetic acidN′-(1-(1-phenyl-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 150)

Compound 150 was prepared according to the procedure described in SchemeI from 1-phenyl-6-trifluoromethyl-1,2-dihydroindol-2-one, triethylorthoacetate, and acetic acid hydrazide.

Example 51

(±)-2-Hydroxypropionic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 151)

Compound 151 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 2-hydroxypropionic acid hydrazide. MW=433,LC-MS (M+1)=433.94.

Example 52

2-Hydroxyacetic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 152)

Compound 152 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 2-hydroxyacetic acid hydrazide. MW=419, LC-MS(M+1)=419.97.

Example 53

3-Ethoxy-3-oxopropionic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 153)

Compound 153 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 3-ethoxy-3-oxopropionic acid hydrazide.MW=475, LC-MS (M+1)=475.91.

Example 54

Aminocarboxylic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 154)

Compound 154 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 1-aminocarboxylic acid hydrazide. MW=404,LC-MS (M+1)=404.93.

Example 55

Acetic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-phenyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 155)

Compound 155 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-phenyl-1,2-dihydroindol-2-one, triethylorthoacetate, and acetic acid hydrazide.

Example 56

2-(4-Hydroxyphenyl)acetic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 156)

Compound 156 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and 4-hydroxyphenylacetic acid hydrazide. ¹H NMR(400 MHz, DMSO-d₆) 11.55 (s, 1H), 10.69 (s, 1H), 9.30 (s, 1H), 7.64 (d,J=8.0, 1H), 7.33 (d, J=8.0, 5H), 7.13 (s, 1H), 7.11 (s, 1H), 7.02 (s,2H), 6.88 (s, 1H), 6.72 (d, J=8.0, 2H), 3.45 (s, 2H), 2.44 (s, 3H), 2.34(s, 6H).

Example 57

Propionic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 157)

Compound 157 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and propionic acid hydrazide. ¹H NMR (400 MHz,DMSO-d₆) 11.50 (s, 1H), 10.49 (s, 1H), 7.68 (d, J=8.0, 1H), 7.36 (d,J=8.0, 3H), 7.13 (s, 1H), 7.04 (s, 2H), 6.89 (s, 1H), 2.45 (s, 3H), 2.34(s, 6H), 2.25 (dd, J=5.0 and 8.0, 2H), 1.10 (dd, J=5.0 and 8.0, 3H).

Example 58

Acetic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 158)

Compound 158 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and acetic acid hydrazide. ¹H NMR (400 MHz,DMSO-d₆) 11.50 (s, 1H), 10.51 (s, 1H), 7.68 (d, J=8.0, 1H), 7.38 (d,J=8.0, 3H), 7.12 (s, 1H), 7.04 (s, 2H), 6.89 (s, 1H), 2.44 (s, 3H), 2.34(s, 6H), 1.99 (s, 3H).

Example 59

Ureidoacetic acidN′-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)-1-phenylmethyl)hydrazide(Compound 159)

Compound 159 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthobenzoate, and 2-(aminoacetylamino)acetic acid hydrazide.MW=523, LC-MS (M+1)=523.97.

Example 60

1-(2-Cyanoethyl)-3-methyl-5-oxo-4,5-dihydro-4-(2-hydroxy-3-(2-(2,6-dimethylphenyl)-ethyl)phenylhydrazono)pyrazole(Compound 160)

Compound 160 was prepared according to the procedure described in SchemeII from 2-hydroxy-3-(2-(2,6-dimethylphenyl)ethyl)aniline, ethylacetoacetate, and 2-cyanoethylhydrazine.

Example 61

1-(3-Hydroxypropyl)-3-methyl-5-oxo-4,5-dihydro-4-(2-hydroxy-3-(2-(2,6-dimethylphenyl)-ethyl)phenylhydrazono)pyrazole(Compound 161)

Compound 161 was prepared according to the procedure described in SchemeII from 2-hydroxy-3-(2-(2,6-dimethylphenyl)ethyl)aniline, ethylacetoacetate, and 3-hydroxypropyl-1-hydrazine.

Example 62

1-(2-Hydroxyethyl)-3-methyl-5-oxo-4,5-dihydro-4-(2-hydroxy-3-(2-(2,5-dimethylphenyl)-ethyl)phenylhydrazono)pyrazole(Compound 162)

Compound 162 was prepared according to the procedure described in SchemeII from 2-hydroxy-3-(2-(2,5-dimethylphenyl)ethyl)aniline, ethylacetoacetate, and 2-hydroxyethyl-1-hydrazine.

Example 63

1-(4-Hydroxybutyl)-3-methyl-5-oxo-4,5-dihydro-4-(2-hydroxy-3-(2(E)-(2-methylphenyl)-ethenyl)phenylhydrazono)pyrazole(Compound 163)

Compound 163 was prepared according to the procedure described in SchemeII from 2-hydroxy-3-(2(E)-(2-methylphenyl)ethenyl)aniline, ethylacetoacetate, and 4-hydroxybutylhydrazine. MW=406, LC-MS (M+1)=407.02.

Example 64

1-(3-Hydroxypropyl)-3-methyl-5-oxo-4,5-dihydro-4-(2-hydroxy-3-(2-(2,5-dimethylphenyl)-ethyl)phenylhydrazono)pyrazole(Compound 164)

Compound 164 was prepared according to the procedure described in SchemeII from 2-hydroxy-3-(2-(2,5-dimethylphenyl)ethyl)aniline, ethylacetoacetate, and 3-hydroxypropyl-1-hydrazine.

Example 65

1-(4-Hydroxybutyl)-3-methyl-5-oxo-4,5-dihydro-4-(2-hydroxy-3-(2-(2,6-dimethylphenyl)-ethynyl)phenylhydrazono)pyrazole(Compound 165)

Compound 165 was prepared according to the procedure described in SchemeII from 2-hydroxy-3-(2-(2,6-dimethylphenyl)ethynyl)aniline, ethylacetoacetate, and 4-hydroxypbutyl-1-hydrazine.

Example 66

1-(2-Cyanoethyl)-3-methyl-5-oxo-4,5-dihydro-4-(2-hydroxy-3-(2-(2,5-dimethylphenyl)-ethyl)phenylhydrazono)pyrazole(Compound 166)

Compound 166 was prepared according to the procedure described in SchemeII from 2-hydroxy-3-(2-(2,5-dimethylphenyl)ethyl)aniline, ethylacetoacetate, and 2-cyanoethyl-1-hydrazine.

Example 67

1-(4-Hydroxybutyl)-3-methyl-5-oxo-4,5-dihydro-4-(2-hydroxy-3-(2-(2-methylphenyl)-ethyl)phenylhydrazono)pyrazole(Compound 167)

Compound 167 was prepared according to the procedure described in SchemeII from 2-hydroxy-3-(2-(2-methylphenyl)ethyl)aniline, ethylacetoacetate, and 4-hydroxybutyl-1-hydrazine.

Example 68

1-Benzyl-3-methyl-5-oxo-4,5-dihydro-4-(2-hydroxy-3-(2-(2,6-dimethylphenyl)-ethyl)phenylhydrazono)pyrazole(Compound 168)

Compound 168 was prepared according to the procedure described in SchemeII from 2-hydroxy-3-(2-(2,6-dimethylphenyl)ethyl)aniline, ethylacetoacetate, and benzylhydrazine.

Example 69

1-(3,5-Dimethylphenyl)-2-oxo-2,3-dihydro-3-(2-hydroxy-4-hydroxymethylphenylhydrazono)-6-trifluoromethylindole(Compound 169)

Compound 169 was prepared according to the procedure similar asdescribed in Scheme II from 2-hydroxy-4-hydroxymethylaniline and1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one.

Example 70

1-(3,5-Dimethylphenyl)-2-oxo-2,3-dihydro-3-(2-hydroxy-3-hydroxymethylphenylhydrazono)-6-trifluoromethylindole(Compound 170)

Compound 170 was prepared according to the procedure similar asdescribed in Scheme II from 2-hydroxy-3-hydroxymethylaniline and1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one.

Example 71

4-(4,5-Dihydro-4-(1-hydroxy-2-naphthylhydrazono)-3-methy-5-oxo-pyrazole)benzoicacid (Compound 171)

Compound 171 was prepared according to the procedure described in SchemeII from 1-hydroxy-2-aminonaphthalene, ethyl acetoacetate, and4-hydroxycarbonylphenylhydrazine.

Example 72

4,5-Dihydro-1-(4-hydroxymethylphenyl)-4-(1-hydroxy-2-naphthylhydrazono)-3-methypyrazol-5-one(Compound 172)

Compound 172 was prepared according to the procedure described in SchemeII from 1-hydroxy-2-aminonaphthalene, ethyl acetoacetate, and4-hydroxymethylphenylhydrazine.

Example 73

4,5-Dihydro-1-(4-(2-cyanoethyl)phenyl)-4-(1-hydroxy-2-naphthylhydrazono)-3-methypyrazol-5-one(Compound 173)

Compound 173 was prepared according to the procedure described in SchemeII from 1-hydroxy-2-aminonaphthalene, ethyl acetoacetate, and4-(2-cyanoethyl)phenylhydrazine.

Example 74

4-Hydroxybutyric acidN′-(1-(5-chloro-1-phenyl-2-oxo-1,2-dihydroindol-3(Z)-ylidene)ethyl)hydrazide(Compound 174)

Compound 174 was prepared according to the procedure described in SchemeI from 1-phenyl-5-chloro-1,2-dihydroindol-2-one, triethyl orthoacetate,and 4-hydroxybutyric acid hydrazide.

Example 75

Methyl2-(1-(1-(3,5-dimethylphenyl)-2-oxo-6-trifluoromethyl-1,2-dihydroindol-3(Z)-ylidene)ethyl)aminoacetate(Compound 175)

Compound 175 was prepared according to the procedure described in SchemeI from 1-(3,5-dimethylphenyl)-6-trifluoromethyl-1,2-dihydroindol-2-one,triethyl orthoacetate, and methyl aminoacetate. MW=418, LC-MS(M+1)=418.96.

Example 76

Acetic acidN′-(1-(1-(3,5-dimethylphenyl)-5-oxo-3-methyl-4,5-dihydropyrazol-4(Z)-ylidene)ethyl)hydrazide(Compound 176)

Compound 176 was prepared according to the procedure similar asdescribed in Scheme I from 1-(3,5-dimethylphenyl)-3-methylpyrazol-5-one,triethyl orthoacetate, and acetic acid hydrazide.

Example 77 Certain Assays

In certain embodiments, assays may be used to determine the level ofGCSFR modulating activity of the compounds of the present embodiments.In certain embodiments, assays containing selectively mutated GCSFR maybe used to determine the interaction of the compounds with the TMdomain. In certain embodiments, assays containing GCSFR from speciesdifferent than human may be used to measure the activity of thecompounds (e.g. mouse or monkey).

Cell Proliferation Assay

In some embodiments, compounds are tested in an in vitro proliferationassay using the cell lines that express GCSFR either endogenously or bystable or transient transfection, and may be dependant upon GCSF fortheir growth. Activity of the compounds on cell proliferation isdetermined by counting the cells or by use of an assay that measures theproduction of ATP as a marker of cell growth.

Reporter Assay

In some embodiments, compounds are tested in a reporter assay using thecell lines that express GCSFR endogenously or by stable or transienttransfection. These cells are transfected, stably or transiently, withthe GCSF-responsive reporter (such as luciferase) and the activity ofthe compounds is determined by measuring the amount of reporter in thecell.

Differentiation Assay

In some embodiments, compounds are tested in purified human CD34+ bonemarrow cells. After addition of the compounds to the cells, the numberof cells expressing markers of hematopoiesis, erythropoiesis,granulopoiesis, thrombopoiesis, or myelopoiesis is measured by flowcytometry, by analyzing expression of genes associated with thesepathways, or by measuring the formation of colonies of cells specificfor these pathways (e.g. erythrocytes or granulocytes). In someembodiments, compounds are tested in bone marrow cells from otherspecies besides human (e.g. mouse or monkey).

Example 78 Luciferase Reporter Assays

Human GCSF Receptor (hGCSFR) Assay

A luciferase reporter assay was used to identify compounds that activatehGCSFR. A human cell line (e.g. MCF-7 breast cancer cells, HepG2hepatoma cells or HEK293 kidney cells) was transiently transfected withan hGCSFR expression plasmid, containing the hGCSFR cDNA downstream ofthe CMV promoter, and a luciferase reporter plasmid, containing aGCSF-responsive element upstream of a minimal thymidine kinase promoter.For the luciferase reporter assay, cells were plated into 96-wellmicrotiter plates in Eagle's Minimal Essential Media (EMEM) containing10% fetal bovine serum (FBS). The following day, the hGCSFR expressionplasmid and the luciferase reporter plasmid were transiently transfectedinto the cells using FuGene6 transfection reagent (Roche, Indianapolis,Ind.), according to manufacturer's specifications. The next day, themedia was replaced with media containing 1% FBS and 10 μM zinc chloride.On the fourth day, media containing recombinant hGCSF (rhGCSF) (at amaximal concentration of 200 ng/ml) or test compound (at a maximalconcentration of 10 μM) were added to the cells in duplicate. After sixhours, the medium was removed from the cells, and the cells were lysedwith a detergent-containing buffer. Luciferase activity was measured incell extracts to determine the level of transcriptional activation.Luciferin-containing buffer was added to each well of the 96-well plateand luciferase activity was measured using a luminometer.

Mutant hGCSFR or Mouse GCSFR (mGCSFR) Assays

A luciferase reporter assay was used to determine the ability ofcompounds to activate mutated hGCSFR, mGCSFR, or mutated mGCSFR. A humancell line (e.g. MCF-7 breast cancer cells, HepG2 hepatoma cells orHEK293 kidney cells) was transiently transfected with one of three GCSFRexpression plasmids: 1) an hGCSFR expression plasmid containing thehGCSFR cDNA in which the nucleotides coding for histidine-627 in thehGCSFR TM domain were changed to code for asparagine that exists asasparagine-602 of the mouse GCSFR TM in a similar position ofhistidine-627; 2) a mGCSFR expression plasmid, containing the mGCSFRcDNA downstream of the CMV promoter; 3) a mGCSFR expression plasmidcontaining the mGCSFR cDNA in which the nucleotides coding forasparagine-602 in the mGCSFR TM domain were changed to code forhistidine. For the luciferase reporter assay, cells were plated into96-well microtiter plates in Eagle's Minimal Essential Media (EMEM)containing 10% fetal bovine serum (FBS). The following day, one of thethree GCSFR expression plasmids and the luciferase reporter plasmid weretransiently transfected into the cells using FuGene6 transfectionreagent (Roche, Indianapolis, Ind.), according to manufacturer'sspecifications. The next day, the media was replaced with mediacontaining 1% FBS and 10 μM zinc chloride. On the fourth day, mediacontaining rhGCSF (at a maximal concentration of 200 ng/ml) or testcompound (at a maximal concentration of 10 μM) were added to the cellsin duplicate. After six hours, the medium was removed from the cells,and the cells were lysed with a detergent-containing buffer. Luciferaseactivity was measured in cell extracts to determine the level oftranscriptional activation. Luciferin-containing buffer was added toeach well of the 96-well plate and luciferase activity was measuredusing a luminometer.

Example 79 Cell Proliferation Assay

A proliferation assay, using the ViaLight™ Plus kit (Cambrex) or theATPlite kit (Roche), has been used to establish the activity ofcompounds on the growth of the cells. The UT-7 human leukemia cell line(Komatsu N, Nakauchi H, Miwa A, Ishihara T, Eguchi M, Moroi M, Okada M,Sat0 Y, Wada H, Yawata Y, Suda T, Miura Y: Establishment andcharacterization of a human leukemic cell line with megakaryocyticfeatures: Dependency on granulocyte-macrophage colony-stimulatingfactor, interleukin 3, or erythropoietin for growth and survival. CancerRes 51:341, 1991) was stably transfected with an hGCSFR expressionplasmid. Stable clones expressing the hGCSFR (UT7-hPG cells) wereidentified and grown and maintained in media containing 10% FBS,rhGM-CSF and G418. For the assay, UT7-hPG cells were starved of rhGM-CSFovernight, and plated in 96-well plates in media containing 10% FBS and10 μM zinc. Media containing rhGCSF (at a maximal concentration of 200ng/ml) or test compound (at a maximal concentration of 10 μM) were addedto the cells in duplicate. After 48 hours, the proliferation of thecells was measured using the ViaLight™ Plus kit according to themanufacturer's protocol. The cells were lysed with adetergent-containing buffer for 10 minutes, and then the ATP monitoringreagent was added to generate luminescent signal. The signal wasmeasured as relative light units in cell extracts to determine the levelof proliferation of the cells.

Example 80 [¹²⁵I]-rhGCSF Competitive Binding Assay

A competitive binding assay was used to determine the activity ofcompounds on the binding of recombinant human GCSF (rhGCSF) to thehGCSFR. For the assay, UT7-hPG cells were starved of rhGM-CSF for 4hours in media containing 2% FBS, and plated in 96-well v-bottom platesin media containing 2% FBS, 10 μM zinc and 0.1% sodium azide. Mediacontaining rhGCSF (at a maximal concentration of 200 ng/ml) or testcompound (at a maximal concentration of 10 μM) were added to the cellsin duplicate. After 1 hour at room temperature, media containing[125I]-labeled rhGCSF (NEX-426, Perkin Elmer) to a final concentrationof 0.05 nM was added to the cells, and the cells were incubated for 2hours at room temperature on a rocking platform. The cells were washedtwice with cold phosphate buffered saline, and the cells were lysed witha triton X-100 containing-buffer. The lysates were transferred toscintillation vials, scintillation fluid added, and the amount ofradiation in the vials (count per minute) measured in a scintillationcounter.

Example 81 Bone Marrow Cell Differentiation Assay

A human bone marrow granulocyte differentiation assay was used todetermine the capacity of rhGCSF or test compounds to induce thedifferentiation of human bone marrow CD34+ cells to CD15-positive(CD15⁺) granulocytes. Purified normal human bone marrow CD34+ cells fromCambrex were used in this assay. The cells were cultured with mediacontaining 10% fetal bovine serum (FBS), 0.5 ng/mL stem cell factor(SCF) with or without 0.1 ng/ml rhGCSF for three days. Cells were washedand plated in 12-well plates in media containing 10% FBS and 0.5 ng/mlSCF. rhGCSF (at a maximal concentration of 100 ng/ml) and test compounds(at a maximal concentration of 1 μM) were added in triplicate, and cellswere cultured an additional 7 days. The cells were stained withanti-CD15 antibody or isotype controls and analyzed by flow cytometry.The activity was measured as percent CD15⁺ cells and presented as thepercentage of maximal rhGCSF response. The EC₅₀ was determined from thedose-response curve of the compound, and efficacy was calculated bycomparison with 100 ng/ml rhGCSF. In addition, the effect of rhGCSF ortest compound on the growth of CD34+ cells was measured using theATPlite™ kit after 7 days of incubation with rhGCSF or compound.

Example 82 A human GCSFR Agonist

GCSF is a cytokine that regulates neutrophilic granulocytes. GCSF actson the homodimeric receptor (GCSFR) to stimulate proliferation ofgranulocytic progenitor cells and induce their survival anddifferentiation into neutrophils. Recombinant human GCSF (rhGCSF) isused successfully to alleviate severe chronic neutropenia andneutropenia induced by chemotherapy or associated with hematopoieticstem cell transplantation. A small molecule oral GCSFR agonist may offera safer and more convenient alternative to the current injectable rhGCSFtherapy. A series of novel non-peptidyl small molecules, exemplified bythe lead Compound X, that selectively activate human GCSFR (hGCSFR)function, and may provide a significant innovation in the treatment ofneutropenia have been discovered.

Methods

Luciferase Reporter Assays: HepG2 or HEK293 cells were transientlytransfected with an expression vector for hGCSFR, hTPOR, hEPOR, mouseGCSFR or mutated receptors and either a STAT3-responsive or aSTAT5-responsive luciferase reporter. Cells were treated with vehicle,rhGCSF (Neupogen®), rhTPO (R&D Systems), rhEPO (Epogen®) or Compound Xfor 6 hours prior to lysis and luciferase measurement.

Generation of hGCSFR Stable Cell Line UTP-hGCSFR: UT7 parental cellswere transfected with an hGCSFR expression vector containing a neomycinresistance gene, and clones were identified by resistance to G418. Asubclone (UTP-hGCSFR) responsive to hGCSF was identified.

Viability Assays: UT7-TPO, UT7-EPO or UTP-hGCSFR cells were treated withvehicle, rhGCSF, rhTPO, rhEPO or Compound X for 48 hours. Viability wasassayed by ATPlight® (PerkinElmer).

Granulocyte Differentiation Assay: CD34-positive human bone marrow cellswere cultured in media supplemented with human stem cell factor (hSCF,R&D Systems), and with vehicle, rhGCSF, or Compound X for 7 days. Thecells were stained with anti-CD15 (BD Biosciences) and analyzed by FACS.

Detection of phospho-STAT3 and phospho-STAT5: Cells cultured withvehicle, rhGCSF or Compound X for various times and phospho-STAT3 andphospho-STAT5 were measured using AlphaScreen® SureFire® assay kits(PerkinElmer).

Radioligand-binding Experiments: UTP-hGCSFR cells were incubated with[125I]rhGCSF (PerkinElmer) in the presence or absence of rhGCSF orCompound X for 2 h at room temperature. Cells were washed twice and theamount of radioactivity measured in a scintillation counter.

Results

Compound X activated STAT3 and STAT5 responsive luciferase reporters incells co-transfected with an hGCSFR expression vector (FIG. 1A and FIG.1B). Compound X did not activate either reporter in the absence oftransfected hGCSFR (not shown). Compound X did not activate aSTAT5-responsive luciferase reporter in HEK293 cells transfected withhTPOR or hEPOR expression vectors (FIG. 2A and FIG. 2B).

Compound X increased the viability of UTP-hGCSFR stable cells (FIG. 3)but did not increase growth of TPO- or EPO-responsive UT-7 cells (datanot shown). Compound X increased the phosphorylation of STAT3 and STAT5in UTP-hGCSFR stable cells (FIG. 3B and FIG. 3C).

Compound X induces the expression of the granulocyte-specific markerCD15 (FUT4) in CD34-positive, human bone marrow cells (BM-HCs) (FIG. 4).Compound X was additive to the effect of 0.1 ng/ml rhGCSF, comparable toa normal serum concentration of GCSF.

The amino acid sequences of the transmembrane domain of GCSFR fromvarious species were compared (FIG. 5). Compound X is active in cellsexpressing monkey, but not mouse, guinea pig or rabbit GCSFR (data notshown). SEQ ID NO.:01 is transmembrane domain of GCSFR from human; SEQID NO.:02 is transmembrane domain of GCSFR from monkey; SEQ ID NO.:03 istransmembrane domain of GCSFR from rabbit; SEQ ID NO.:04 istransmembrane domain of GCSFR from guinea pig; SEQ ID NO.:05 istransmembrane domain of GCSFR from mouse.

Mutant receptors were created by replacing H627 in hGCSFR with N presentat a similar location in the mouse GCSFR (N602), and by replacing N602in the mouse GCSFR with H. Compound X is not active in cells expressingthe mGCSFR or hGCSFR-H627N (FIG. 6A). Compound X is active, however, onmGCSFR-N602H (FIG. 6B). Mean (triplicates) ±SD.

Compound X did not displace [125I]rhGCSF binding to UTP-hGCSFR cells,however binding of [125I]rhGCSF was augmented in aconcentration-dependent manner (FIG. 7).

Discussion

A novel small molecule human GCSFR agonist that activates GCSFR functionwas discovered. Compound X was found to activate the GCSFR/JAK/STATsignal transduction pathway and increased the viability of cellscontaining hGCSFR. Compound X promoted the differentiation of bonemarrow cells into granulocytes, increasing expression of CD15. CompoundX is dependent on the expression of hGCSFR and a histidine in thereceptor transmembrane domain was needed for activity, similar to whathas been found for small-molecule human TPOR agonists. Compound Xincreased the binding of GCSF in a manner consistent with allostericreceptor modulation.

These data demonstrate that Compound X is a novel small-moleculeselective hGCSFR agonist that activates the receptor in a mannerdistinct from GCSF and similar to the mechanism of small-molecule hTPORagonists. Further optimization of the Compound X chemical series shouldyield a new generation of orally-available molecules to treatneutropenia with improved safety and convenience compared to currentinjectable rhGCSF.

The term “comprising” as used herein is synonymous with “including,”“containing,” or “characterized by,” and is inclusive or open-ended anddoes not exclude additional, unrecited elements or method steps.

All numbers expressing quantities of ingredients, reaction conditions,and so forth used in the specification are to be understood as beingmodified in all instances by the term “about.” Accordingly, unlessindicated to the contrary, the numerical parameters set forth herein areapproximations that may vary depending upon the desired propertiessought to be obtained. At the very least, and not as an attempt to limitthe application of the doctrine of equivalents to the scope of anyclaims in any application claiming priority to the present application,each numerical parameter should be construed in light of the number ofsignificant digits and ordinary rounding approaches.

The above description discloses several methods and materials of thepresent invention. This invention is susceptible to modifications in themethods and materials, as well as alterations in the fabrication methodsand equipment. Such modifications will become apparent to those skilledin the art from a consideration of this disclosure or practice of theinvention disclosed herein. Consequently, it is not intended that thisinvention be limited to the specific embodiments disclosed herein, butthat it cover all modifications and alternatives coming within the truescope and spirit of the invention.

All references cited herein, including but not limited to published andunpublished applications, patents, and literature references, areincorporated herein by reference in their entirety and are hereby made apart of this specification. To the extent publications and patents orpatent applications incorporated by reference contradict the disclosurecontained in the specification, the specification is intended tosupersede and/or take precedence over any such contradictory material.

What is claimed is:
 1. A compound of Formula (I), (II), or (IV):

a tautomer thereof, or a pharmaceutically acceptable salt thereof,wherein: R¹ is selected from hydrogen, OR⁶, NO₂, CN, NR⁶R⁷, CO₂R⁶,C(═O)NR⁶R⁷, SO₃R⁶, SO₂NR⁶R⁸, an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₂-C₆ alkenyl, an optionally substituted C₂-C₆alkynyl, an optionally substituted C₁-C₆ heteroalkyl, an optionallysubstituted C₃-C₆ cycloalkyl, an optionally substituted C₃-C₆cycloalkenyl, an optionally substituted C₂-C₆ heterocyclyl, anoptionally substituted arylalkyl, an optionally substituted aryl, and anoptionally substituted heteroaryl; R² and R³ are independently selectedfrom hydrogen, an optionally substituted C₁-C₆ alkyl, an optionallysubstituted C₂-C₆ alkenyl, an optionally substituted C₂-C₆ alkynyl, anoptionally substituted C₁-C₆ heteroalkyl, an optionally substitutedC₃-C₈ cycloalkyl, an optionally substituted C₃-C₈ cycloalkenyl, anoptionally substituted C₁-C₆ heterocycle, an optionally substitutedaryl, and an optionally substituted heteroaryl; R⁴ is selected fromhydrogen, an optionally substituted C₁-C₆ alkyl, an optionallysubstituted C₂-C₆ alkenyl, an optionally substituted C₂-C₆ alkynyl, anoptionally substituted C₁-C₆ heteroalkyl, an optionally substitutedC₃-C₈ cycloalkyl, an optionally substituted C₃-C₈ cycloalkenyl, anoptionally substituted C₁-C₆ heterocycle, an optionally substitutedaryl, an optionally substituted heteroaryl, an optionally substitutedarylalkyl, an optionally substituted arylalkenyl, an optionallysubstituted arylalkynyl, and an optionally substituted heteroarylalkyl;R⁵ is selected from hydrogen, halogen, NO₂, CN, CF₃, OR⁶, CO₂R⁶,C(═O)NR⁶R⁷, SO₃R⁶, and SO₂NR⁶R⁸, an optionally substituted aryl, anoptionally substituted C₁-C₆ alkyl, and an optionally substituted C₁-C₆heteroalkyl; R⁶ is selected from hydrogen, an optionally substitutedC₁-C₆ alkyl, a C₁-C₆ heteroalkyl, an optionally substituted aryl, and anoptionally substituted heteroaryl; R⁷ is selected from hydrogen,C(═O)R⁸, C(═O)NHR⁸, an optionally substituted C₁-C₆ alkyl, and anoptionally substituted C₁-C₆ heteroalkyl; or —NR⁶R⁷ is an optionallysubstituted non-aromatic heterocycle linked through a ring nitrogen; R⁸is selected from hydrogen, an optionally substituted C₁-C₆ alkyl, and anoptionally substituted C₁-C₆ heteroalkyl; R⁹ is selected from hydrogen,an optionally substituted C₁-C₆ alkyl, an optionally substituted C₂-C₆alkenyl, an optionally substituted C₂-C₆ alkynyl, an optionallysubstituted C₁-C₆ heteroalkyl, an optionally substituted C₃-C₈cycloalkyl, an optionally substituted C₃-C₈ cycloalkenyl, an optionallysubstituted C₁-C₆ heterocycle, an optionally substituted heteroaryl, anoptionally substituted arylalkyl, an optionally substituted arylalkenyl,an optionally substituted arylalkynyl, an optionally substitutedheteroarylalkyl, an optionally substituted heteroarylalkenyl, and anoptionally substituted heteroarylalkynyl; Q is selected from the groupconsisting of NR⁶, an optionally substituted C₁-C₆ alkyl, an optionallysubstituted C₃-C₈ cycloalkyl, an optionally substituted C₁-C₆heteroalkyl, and an optionally substituted non-aromatic heterocycle; L¹is selected from NH and CHR²; W is selected from O (oxygen) and NH; X isN (nitrogen) or CR²; Y is selected from an optionally substituted C₁-C₆alkyl, C₂-C₆ alkenyl, an optionally substituted C₁-C₆ heteroalkyl, anoptionally substituted C₁-C₆ heteroalkenyl, an optionally substitutedphenylalkenyl, and an optionally substituted heterocyclealkenyl; Z is O(oxygen) or S (sulfur); n is 1, 2 or 3; and with the proviso that if R²is methyl, R⁴ is phenyl, L¹ is NH, and Q is N-Ph-R¹ in Formula I andIII, R¹ of Formula I and III is not selected from the group of halogen,alkyl, substituted alkyl, carboxylic acid, and carboxylic esters.
 2. Thecompound of claim 1 having the structure of Formula (Ia):

a tautomer thereof, or a pharmaceutically acceptable salt thereof,wherein: R¹ is selected from hydrogen, an optionally substituted C₁-C₆alkyl, an optionally substituted C₃-C₆ cycloalkyl, an optionallysubstituted C₃-C₆ cycloalkenyl, an optionally substituted C₂-C₆heterocyclyl, an optionally substituted aryl, and an optionallysubstituted heteroaryl; R² and R³ are independently selected fromhydrogen, an optionally substituted C₁-C₆ alkyl, an optionallysubstituted C₃-C₈ cycloalkyl, an optionally substituted C₁-C₆heterocycle, an optionally substituted aryl, and an optionallysubstituted heteroaryl; R⁴ is selected from hydrogen, an optionallysubstituted C₁-C₆ alkyl, an optionally substituted C₃-C₈ cycloalkyl, anoptionally substituted C₃-C₈ cycloalkenyl, an optionally substitutedC₁-C₆ heterocycle, an optionally substituted aryl, and an optionallysubstituted heteroaryl; Q is selected from an optionally substitutedC₁-C₆ alkyl, an optionally substituted C₃-C₈ cycloalkyl, and anoptionally substituted non-aromatic heterocycle; and Z is O (oxygen) orS (sulfur).
 3. The compound of claim 2, wherein: R¹ is selected fromhydrogen, and an optionally substituted C₁-C₆ alkyl; R² and R³ areindependently selected from hydrogen, and an optionally substitutedC₁-C₆ alkyl; R⁴ is selected from hydrogen, an optionally substitutedC₁-C₆ alkyl, an optionally substituted aryl, and an optionallysubstituted heteroaryl; Q is selected from an optionally substitutedC₁-C₆ alkyl, and an optionally substituted C₃-C₈ cycloalkyl; and Z is O(oxygen).
 4. The compound of claim 3, wherein: R¹ is selected fromhydrogen, and an optionally substituted C₁-C₃ alkyl; R² and R³ areindependently selected from hydrogen, and an optionally substitutedC₁-C₃ alkyl; R⁴ is selected from optionally substituted C₁-C₆ alkyl, andan optionally substituted aryl; and Q is selected from an optionallysubstituted C₁-C₃ alkyl, and an optionally substituted C₃-C₃ cycloalkyl.5. The compound of claim 4, wherein: R¹ is hydrogen; R² and R³ areindependently an optionally substituted C₁-C₃ alkyl; R⁴ is an optionallysubstituted phenyl; and Q is an optionally substituted C₁-C₃ alkyl. 6.The compound of claim 1 having the structure of Formula (IIa) or (IIb):

a tautomer thereof, or a pharmaceutically acceptable salt thereof,wherein: R³ is selected from hydrogen, an optionally substituted C₁-C₆alkyl, an optionally substituted C₃-C₈ cycloalkyl, an optionallysubstituted C₁-C₆ heterocycle, an optionally substituted aryl, and anoptionally substituted heteroaryl; R⁴ is selected from an optionallysubstituted C₁-C₆ alkyl, an optionally substituted C₃-C₈ cycloalkyl, anoptionally substituted C₁-C₆ heterocyclyl, an optionally substitutedaryl, an optionally substituted heteroaryl, an optionally substitutedarylalkyl, and an optionally substituted heteroarylalkyl; and R⁹ isselected from hydrogen, an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₃-C₈ cycloalkyl, an optionally substituted C₁-C₆heterocyclyl, an optionally substituted heteroaryl, an optionallysubstituted arylalkyl, and an optionally substituted heteroarylalkyl. 7.The compound of claim 6, wherein: R³ is selected from hydrogen, and anoptionally substituted C₁-C₃ alkyl; R⁴ is selected from an optionallysubstituted C₁-C₆ alkyl, an optionally substituted C₃-C₈ cycloalkyl, andan optionally substituted aryl; and R⁹ is selected from an optionallysubstituted C₁-C₆ alkyl, an optionally substituted arylalkyl, anoptionally substituted arylalkenyl, an optionally substitutedarylalkynyl, and an optionally substituted heteroarylalkyl.
 8. Thecompound of claim 7, wherein: R³ is an optionally substituted C₁-C₃alkyl; R⁴ is selected from an optionally substituted C₁-C₄ alkyl, and anoptionally substituted aryl; and R⁹ is selected from an optionallysubstituted C₁-C₃ alkyl, an optionally substituted arylalkyl, anoptionally substituted arylalkenyl, and an optionally substitutedarylalkynyl.
 9. The compound of claim 1 having the structure of Formula(IVa) or (IVb):

a tautomer thereof, or a pharmaceutically acceptable salt thereof,wherein: R⁴ is selected from an optionally substituted C₁-C₆ alkyl, anoptionally substituted C₃-C₈ cycloalkyl, an optionally substituted C₁-C₆heterocyclyl, an optionally substituted aryl, an optionally substitutedheteroaryl, an optionally substituted arylalkyl, and an optionallysubstituted heteroarylalkyl; R⁵ is selected from halogen, CN, CF₃, OR⁶,an optionally substituted aryl, and an optionally substituted C₁-C₆alkyl; R⁶ is selected from hydrogen, an optionally substituted C₁-C₆alkyl, an optionally substituted aryl, and an optionally substitutedheteroaryl; and R⁹ is selected from hydrogen, an optionally substitutedC₁-C₆ alkyl, an optionally substituted C₃-C₈ cycloalkyl, an optionallysubstituted C₁-C₆ heterocyclyl, an optionally substituted heteroaryl, anoptionally substituted arylalkyl, and an optionally substitutedheteroarylalkyl.
 10. The compound of claim 9, wherein: R⁴ is selectedfrom an optionally substituted C₁-C₆ alkyl, an optionally substitutedC₃-C₈ cycloalkyl, and an optionally substituted aryl; R⁵ is selectedfrom chloro, CN, CF₃, OR⁶, an optionally substituted aryl, and anoptionally substituted C₁-C₆ alkyl; R⁶ is selected from hydrogen, and anoptionally substituted C₁-C₃ alkyl; R⁹ is selected from an optionallysubstituted C₁-C₆ alkyl, an optionally substituted arylalkyl, anoptionally substituted arylalkenyl, an optionally substitutedarylalkynyl, and an optionally substituted heteroarylalkyl, and n is 1,or
 2. 11. The compound of claim 10, wherein: R⁴ is selected from anoptionally substituted C₁-C₄ alkyl, and an optionally substituted aryl;R⁵ is selected from chloro, CN, CF₃, and an optionally substituted C₁-C₃alkyl; R⁹ is selected from an optionally substituted C₁-C₃ alkyl, anoptionally substituted arylalkyl, an optionally substituted arylalkenyl,and an optionally substituted arylalkynyl; and n is
 1. 12. Apharmaceutical composition comprising a compound of claim 1 and apharmaceutically acceptable excipient.
 13. A method of treating ahematopoietic or neurological disorder comprising administering aneffective amount of a compound of claim 1 to a subject in need thereof,wherein the compound is a Formula (I) or (IV).
 14. The method of claim13, wherein the disorder is selected from the group consisting ofgranulocytopenia, neutropenia, amyotrophic lateral sclerosis, multiplesclerosis, multiple dystrophy, and spinal cord injury.
 15. The method ofclaim 13, wherein the compound is administered in combination with anadditional therapeutic regimen selected from the group consisting ofchemotherapy, bone marrow transplantation, and radiation therapy
 16. Amethod of treating a hematopoietic or neurological disorder comprisingadministering an effective amount of a compound of Formula (V) or (VI)to a subject in need thereof, wherein Formula (V) or (VI):having thestructure:

or pharmaceutically acceptable salt thereof, wherein: R¹⁰ is selectedfrom an optionally substituted C₁-C₁₂ alkyl, an optionally substitutedC₁-C₁₂ heteroalkyl, an optionally substituted C₂-C₁₂ heteroalkenyl, anoptionally substituted C₂-C₁₂ heteroalkynyl, an optionally substitutedC₁-C₁₂ cycloalkyl; an optionally substituted C₁-C₆ heterocycloalkyl, anoptionally substituted heteroarylalkyl, an optionally substitutedarylheteroalkyl, an optionally substituted heteroarylheteroalkyl, OR¹⁴,SR¹⁴, and NR⁸R¹⁴; R₁₁ is selected from hydrogen, halogen, an optionallysubstituted C₁-C₆ alkyl, an optionally substituted C₁-C₆ heteroalkyl, anoptionally substituted C₃-C₈ cycloalkyl, and an optionally substitutedC₁-C₆ heterocycle; R₁₂ is selected from an optionally substituted C₁-C₆heteroalkyl, an optionally substituted C₃-C₈ cycloalkyl, an optionallysubstituted C₁-C₆ heterocycle, an optionally substituted aryl, and anoptionally substituted heteroaryl; R₁₃ is selected from hydrogen,halogen, an optionally substituted C₁-C₈ alkyl, an optionallysubstituted C₁-C₈ heteroalkyl, an optionally substituted C₂-C₈heteroalkenyl, an optionally substituted C₃-C₈ cycloalkyl, an optionallysubstituted C₁-C₆ heterocycloalkyl, OR¹⁴, SR¹⁴, and NR⁸R¹⁴; R₁₄ isselected from an optionally substituted aryl and an optionallysubstituted heteroaryl; V is selected from CH═CH, N═CH, CH═N, NH, O(oxygen), and S (sulfur); and n is 1,2 or
 3. 17. The method of claim 16,wherein the disorder is selected from the group consisting ofgranulocytopenia, neutropenia, amyotrophic lateral sclerosis, multiplesclerosis, multiple dystrophy, and spinal cord injury.
 18. The method ofclaim 16, wherein the compound is administered in combination with anadditional therapeutic regimen selected from the group consisting ofchemotherapy, bone marrow transplantation, and radiation therapy. 19.The compound of claim 1 having the structure:

a tautomer thereof, or pharmaceutically acceptable salt thereof.